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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2, 4 and 6) using 5 μg of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230). The immunoprecipitated proteins were subsequently analysed by Western blot. The results obtained with the Cas9 antibody are shown in lane 3 and 4. The negative control (IP with beads only) is shown in lane 5 and 6, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/Cas9 antibody (cat. No. C15200230) diluted 1:400 in blocking solution at 4°C o/n, followed by incubation with an anti-mouse secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259), diluited 1:10,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong> <br /> Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310259) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310260), diluited 1:10,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IP.jpg" alt="IP figure 2" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310260). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IF.jpg" alt="IF figure 3" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Diagenode is pleased to offer the <strong>first antibodies</strong> directed against the <strong>CRISPR/Cas9</strong> nuclease from <strong><em>Staphylococcus aureus</em></strong>.</p>
<h3><em>S. aureus</em> Cas9 monoclonal antibody</h3>
<ul>
<li>Excellent results in <strong>WB</strong>,<strong> IP</strong> and<strong> IF</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-WB.jpg" alt="WB figure 1" /></p>
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<p><small> <strong>Western blot analysis using the Diagenode monoclonal antibody directed against <em>S. aureus</em> Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with <em>S. aureus</em> Cas 9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crispr-cas9-monoclonal-antibody" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IF.jpg" alt="IF figure 1" /></p>
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<div class="small-7 columns">
<p><small> <strong>Immunofluorescence using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (N-terminal)</strong><br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the <em>S. aureus</em> CRISPR/Cas9 antibody (cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>C-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310259-IP.jpg" alt="IP figure 1" width="400" height="343" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>IP using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 µg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 µg) is shown in lane 1 and 2.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IP</th>
<th>IF</th>
<th>Antibody raised against</th>
<th>Specificity</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="/p/s-aureus-crispr-cas9-monoclonal-antibody"><em>S. aureus</em> CRISPR/Cas9 monoclonal antibody</a></td>
<td>+++</td>
<td>+++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>C-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
</tbody>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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'description' => '<p>Previous studies from others and us have demonstrated that CRISPR genome editing could offer a promising therapeutic strategy to restore dystrophin expression and function in the skeletal muscle and heart of Duchenne muscular dystrophy (DMD) mouse models. However, the long-term efficacy and safety of CRISPR genome-editing therapy for DMD has not been well established. We packaged both SaCas9 and guide RNA (gRNA) together into one AAVrh.74 vector, injected two such vectors (targeting intron 20 and intron 23, respectively) into mdx pups at day 3 and evaluated the mice at 19 months. We found that AAVrh.74-mediated life-long CRISPR genome editing in mdx mice restored dystrophin expression and improved cardiac function without inducing serious adverse effects. PCR analysis and targeted deep sequencing showed that the DSBs were mainly repaired by the precise ligation of the two cut sites. Serological and histological examination of major vital organs did not reveal any signs of tumor development or other deleterious defects arising from CRISPR genome editing. These results support that in vivo CRISPR genome editing could be developed as a safe therapeutic treatment for DMD and potentially other diseases.</p>',
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'description' => '<p>Duchenne muscular dystrophy (DMD) is a monogenic disorder and a candidate for therapeutic genome editing. There have been several recent reports of genome editing in preclinical models of Duchenne muscular dystrophy, however, the long-term persistence and safety of these genome editing approaches have not been addressed. Here we show that genome editing and dystrophin protein restoration is sustained in the mdx mouse model of Duchenne muscular dystrophy for 1 year after a single intravenous administration of an adeno-associated virus that encodes CRISPR (AAV-CRISPR). We also show that AAV-CRISPR is immunogenic when administered to adult mice; however, humoral and cellular immune responses can be avoided by treating neonatal mice. Additionally, we describe unintended genome and transcript alterations induced by AAV-CRISPR that should be considered for the development of AAV-CRISPR as a therapeutic approach. This study shows the potential of AAV-CRISPR for permanent genome corrections and highlights aspects of host response and alternative genome editing outcomes that require further study.</p>',
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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'isotype' => '',
'lot' => '10µg/50µg: 001 // 100µg: 002',
'concentration' => '2 μg/μl',
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'type' => 'Monoclonal',
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'classification' => 'Classic',
'application_table' => '<table>
<thead>
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<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
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<td>Western Blotting</td>
<td>1:4,000</td>
<td>Fig 1</td>
</tr>
<tr>
<td>Immunoprecipitation</td>
<td>5 μg/IP</td>
<td>Fig 2</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:400</td>
<td>Fig 3</td>
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'name' => '<i>S. aureus</i> CRISPR/Cas9 Antibody',
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'label1' => 'Validation data',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-WB.jpg" alt="CRISPR/Cas9 Antibody validated in WB " caption="false" width="284" height="289" /></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-IP.jpg" alt="CRISPR/Cas9 Antibody validated in IP" caption="false" width="284" height="177" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 2. IP using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2, 4 and 6) using 5 μg of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230). The immunoprecipitated proteins were subsequently analysed by Western blot. The results obtained with the Cas9 antibody are shown in lane 3 and 4. The negative control (IP with beads only) is shown in lane 5 and 6, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-IF.jpg" alt="CRISPR/Cas9 Antibody validated in IF" caption="false" width="284" height="91" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/Cas9 antibody (cat. No. C15200230) diluted 1:400 in blocking solution at 4°C o/n, followed by incubation with an anti-mouse secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259), diluited 1:10,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong> <br /> Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310259) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310260), diluited 1:10,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310260). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'name' => '<i>S. aureus</i> CRISPR/Cas9 antibodies',
'description' => '<p>Diagenode is pleased to offer the <strong>first antibodies</strong> directed against the <strong>CRISPR/Cas9</strong> nuclease from <strong><em>Staphylococcus aureus</em></strong>.</p>
<h3><em>S. aureus</em> Cas9 monoclonal antibody</h3>
<ul>
<li>Excellent results in <strong>WB</strong>,<strong> IP</strong> and<strong> IF</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-WB.jpg" alt="WB figure 1" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Western blot analysis using the Diagenode monoclonal antibody directed against <em>S. aureus</em> Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with <em>S. aureus</em> Cas 9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crispr-cas9-monoclonal-antibody" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IF.jpg" alt="IF figure 1" /></p>
</div>
<div class="small-7 columns">
<p><small> <strong>Immunofluorescence using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (N-terminal)</strong><br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the <em>S. aureus</em> CRISPR/Cas9 antibody (cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>C-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310259-IP.jpg" alt="IP figure 1" width="400" height="343" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>IP using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 µg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 µg) is shown in lane 1 and 2.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IP</th>
<th>IF</th>
<th>Antibody raised against</th>
<th>Specificity</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="/p/s-aureus-crispr-cas9-monoclonal-antibody"><em>S. aureus</em> CRISPR/Cas9 monoclonal antibody</a></td>
<td>+++</td>
<td>+++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>C-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
</tbody>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2, 4 and 6) using 5 μg of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230). The immunoprecipitated proteins were subsequently analysed by Western blot. The results obtained with the Cas9 antibody are shown in lane 3 and 4. The negative control (IP with beads only) is shown in lane 5 and 6, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/Cas9 antibody (cat. No. C15200230) diluted 1:400 in blocking solution at 4°C o/n, followed by incubation with an anti-mouse secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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'description' => '<p>The CRISPR/Cas9 technology is delivering superior genetic models for fundamental disease research, drug screening, therapy development, rapid diagnostics, and transcriptional modulation. Although CRISPR/Cas9 enables rapid genome editing, several aspects affect its efficiency and specificity including guide RNA design, delivery methods, and off-targets effects. Diagenode has developed strategies to overcome these common pitfalls and has optimized CRISPR/Cas9 genome editing specificity</p>',
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/Cas9 antibody (cat. No. C15200230) diluted 1:400 in blocking solution at 4°C o/n, followed by incubation with an anti-mouse secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259), diluited 1:10,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong> <br /> Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310259) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310260). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IF.jpg" alt="IF figure 3" /></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
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'description' => '<p>Diagenode is pleased to offer the <strong>first antibodies</strong> directed against the <strong>CRISPR/Cas9</strong> nuclease from <strong><em>Staphylococcus aureus</em></strong>.</p>
<h3><em>S. aureus</em> Cas9 monoclonal antibody</h3>
<ul>
<li>Excellent results in <strong>WB</strong>,<strong> IP</strong> and<strong> IF</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-WB.jpg" alt="WB figure 1" /></p>
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<p><small> <strong>Western blot analysis using the Diagenode monoclonal antibody directed against <em>S. aureus</em> Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with <em>S. aureus</em> Cas 9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crispr-cas9-monoclonal-antibody" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IF.jpg" alt="IF figure 1" /></p>
</div>
<div class="small-7 columns">
<p><small> <strong>Immunofluorescence using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (N-terminal)</strong><br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the <em>S. aureus</em> CRISPR/Cas9 antibody (cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>C-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310259-IP.jpg" alt="IP figure 1" width="400" height="343" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>IP using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 µg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 µg) is shown in lane 1 and 2.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IP</th>
<th>IF</th>
<th>Antibody raised against</th>
<th>Specificity</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="/p/s-aureus-crispr-cas9-monoclonal-antibody"><em>S. aureus</em> CRISPR/Cas9 monoclonal antibody</a></td>
<td>+++</td>
<td>+++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>C-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
</tbody>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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'description' => '<p>Previous studies from others and us have demonstrated that CRISPR genome editing could offer a promising therapeutic strategy to restore dystrophin expression and function in the skeletal muscle and heart of Duchenne muscular dystrophy (DMD) mouse models. However, the long-term efficacy and safety of CRISPR genome-editing therapy for DMD has not been well established. We packaged both SaCas9 and guide RNA (gRNA) together into one AAVrh.74 vector, injected two such vectors (targeting intron 20 and intron 23, respectively) into mdx pups at day 3 and evaluated the mice at 19 months. We found that AAVrh.74-mediated life-long CRISPR genome editing in mdx mice restored dystrophin expression and improved cardiac function without inducing serious adverse effects. PCR analysis and targeted deep sequencing showed that the DSBs were mainly repaired by the precise ligation of the two cut sites. Serological and histological examination of major vital organs did not reveal any signs of tumor development or other deleterious defects arising from CRISPR genome editing. These results support that in vivo CRISPR genome editing could be developed as a safe therapeutic treatment for DMD and potentially other diseases.</p>',
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2, 4 and 6) using 5 μg of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230). The immunoprecipitated proteins were subsequently analysed by Western blot. The results obtained with the Cas9 antibody are shown in lane 3 and 4. The negative control (IP with beads only) is shown in lane 5 and 6, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<td>Western Blotting</td>
<td>1:4,000</td>
<td>Fig 1</td>
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<tr>
<td>Immunoprecipitation</td>
<td>5 μg/IP</td>
<td>Fig 2</td>
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2, 4 and 6) using 5 μg of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230). The immunoprecipitated proteins were subsequently analysed by Western blot. The results obtained with the Cas9 antibody are shown in lane 3 and 4. The negative control (IP with beads only) is shown in lane 5 and 6, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/Cas9 antibody (cat. No. C15200230) diluted 1:400 in blocking solution at 4°C o/n, followed by incubation with an anti-mouse secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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'info2' => '<p>CRISPR systems are adaptable immune mechanisms which are present in many bacteria to protect themselves from foreign nucleic acids, such as viruses, transposable elements or plasmids. The CRISPR/Cas9 (CRISPR-associated protein 9 nuclease) system from S. pyogenes was the first to be adapted for inducing sequence-specific double stranded breaks and targeted genome editing. This system is unique and flexible due to its dependence on RNA as the moiety that targets the nuclease to a desired DNA sequence and can be used to induce indel mutations, specific sequence replacements or insertions and large deletions or genomic rearrangements at any desired location in the genome. In addition, Cas9 can also be used to mediate upregulation of specific endogenous genes or to alter histone modifications or DNA methylation, Recently, the CRISPR/Cas9 from S. aureus (UniProtKB/Swiss-Prot entry J7RUA5) was also shown to be suitable for humane genome editing. The S. aureus CRISPR/Cas9 has the advantage that it’s smaller and therefore easier to transfect cells with, whereas the efficiency and specificity are similar.</p>',
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259), diluited 1:10,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15310260). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode antibody directed against S. aureus CRISPR/Cas9 (N-terminal)</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/ Cas9 antibody (Cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'name' => '<i>S. aureus</i> CRISPR/Cas9 antibodies',
'description' => '<p>Diagenode is pleased to offer the <strong>first antibodies</strong> directed against the <strong>CRISPR/Cas9</strong> nuclease from <strong><em>Staphylococcus aureus</em></strong>.</p>
<h3><em>S. aureus</em> Cas9 monoclonal antibody</h3>
<ul>
<li>Excellent results in <strong>WB</strong>,<strong> IP</strong> and<strong> IF</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200230-WB.jpg" alt="WB figure 1" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Western blot analysis using the Diagenode monoclonal antibody directed against <em>S. aureus</em> Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with <em>S. aureus</em> Cas 9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crispr-cas9-monoclonal-antibody" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>N-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310260-IF.jpg" alt="IF figure 1" /></p>
</div>
<div class="small-7 columns">
<p><small> <strong>Immunofluorescence using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (N-terminal)</strong><br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the <em>S. aureus</em> CRISPR/Cas9 antibody (cat. No. C15310260) diluted 1:1,000 in blocking solution at 4°C o/n, followed by incubation with an anti-rabbit secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</h3>
<ul>
<li>Validated in <strong>WB</strong>, <strong>IF</strong> and <strong>IP</strong></li>
<li>Raised against <strong>C-terminus</strong> of Cas9 nuclease</li>
</ul>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310259-IP.jpg" alt="IP figure 1" width="400" height="343" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>IP using the Diagenode antibody directed against <em>S. aureus</em> CRISPR/Cas9 (C-terminal)</strong><br />IP was performed on whole cell extracts (200 µg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310259). The immunoprecipitated proteins were subsequently analysed by Western blot with the monoclonal CRISPR/Cas9 antibody (C15200230). Lane 3 and 4 show the result of the IP, the input (10 µg) is shown in lane 1 and 2.</small></p>
<div class="small-12 medium-12 large-12 columns text-right"><a href="../p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal" class="tiny details button radius">Learn more</a></div>
</div>
</div>
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IP</th>
<th>IF</th>
<th>Antibody raised against</th>
<th>Specificity</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="/p/s-aureus-crispr-cas9-monoclonal-antibody"><em>S. aureus</em> CRISPR/Cas9 monoclonal antibody</a></td>
<td>+++</td>
<td>+++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-n-terminal-sample-size"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (N-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>N-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
<tr>
<td><a href="/p/s-aureus-crisprcas9-polyclonal-antibody-c-terminal"><em>S. aureus</em> CRISPR/Cas9 polyclonal antibody (C-terminal)</a></td>
<td>++</td>
<td>++</td>
<td>+++</td>
<td>C-terminus of Cas9 nuclease</td>
<td><em>Staphylococcus aureus</em></td>
</tr>
</tbody>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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'description' => '<p>Previous studies from others and us have demonstrated that CRISPR genome editing could offer a promising therapeutic strategy to restore dystrophin expression and function in the skeletal muscle and heart of Duchenne muscular dystrophy (DMD) mouse models. However, the long-term efficacy and safety of CRISPR genome-editing therapy for DMD has not been well established. We packaged both SaCas9 and guide RNA (gRNA) together into one AAVrh.74 vector, injected two such vectors (targeting intron 20 and intron 23, respectively) into mdx pups at day 3 and evaluated the mice at 19 months. We found that AAVrh.74-mediated life-long CRISPR genome editing in mdx mice restored dystrophin expression and improved cardiac function without inducing serious adverse effects. PCR analysis and targeted deep sequencing showed that the DSBs were mainly repaired by the precise ligation of the two cut sites. Serological and histological examination of major vital organs did not reveal any signs of tumor development or other deleterious defects arising from CRISPR genome editing. These results support that in vivo CRISPR genome editing could be developed as a safe therapeutic treatment for DMD and potentially other diseases.</p>',
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'description' => '<p>Duchenne muscular dystrophy (DMD) is a monogenic disorder and a candidate for therapeutic genome editing. There have been several recent reports of genome editing in preclinical models of Duchenne muscular dystrophy, however, the long-term persistence and safety of these genome editing approaches have not been addressed. Here we show that genome editing and dystrophin protein restoration is sustained in the mdx mouse model of Duchenne muscular dystrophy for 1 year after a single intravenous administration of an adeno-associated virus that encodes CRISPR (AAV-CRISPR). We also show that AAV-CRISPR is immunogenic when administered to adult mice; however, humoral and cellular immune responses can be avoided by treating neonatal mice. Additionally, we describe unintended genome and transcript alterations induced by AAV-CRISPR that should be considered for the development of AAV-CRISPR as a therapeutic approach. This study shows the potential of AAV-CRISPR for permanent genome corrections and highlights aspects of host response and alternative genome editing outcomes that require further study.</p>',
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<p><small> <strong>Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against S. aureus CRISPR/ Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells (lane 1), HEK293 cells transfected with S. aureus CRISPR/Cas9 (lane 2) and HeLa cells transfected with S. pyogenes Cas9 (lane 3) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230), diluited 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.</small></p>
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<p><small><strong>Figure 2. IP using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong><br />IP was performed on whole cell extracts (200 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2, 4 and 6) using 5 μg of the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200230). The immunoprecipitated proteins were subsequently analysed by Western blot. The results obtained with the Cas9 antibody are shown in lane 3 and 4. The negative control (IP with beads only) is shown in lane 5 and 6, the input (10 μg) is shown in lane 1 and 2.</small></p>
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<p><small> <strong>Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against S. aureus CRISPR/Cas9</strong> <br />Transiently transfected U2OS cells expressing SaCas9-T2A-GFP were fixed with 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the S. aureus CRISPR/Cas9 antibody (cat. No. C15200230) diluted 1:400 in blocking solution at 4°C o/n, followed by incubation with an anti-mouse secondary antibody coupled to DyLight594 for 1 h at RT (left figure). Nuclei were counter-stained with Hoechst 33342 (right). The middle figure shows IF with an anti-GFP antibody.</small></p>
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'name' => ' Accurate QC to optimize CRISPR/Cas9 genome editing specificity',
'description' => '<p>The CRISPR/Cas9 technology is delivering superior genetic models for fundamental disease research, drug screening, therapy development, rapid diagnostics, and transcriptional modulation. Although CRISPR/Cas9 enables rapid genome editing, several aspects affect its efficiency and specificity including guide RNA design, delivery methods, and off-targets effects. Diagenode has developed strategies to overcome these common pitfalls and has optimized CRISPR/Cas9 genome editing specificity</p>',
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'description' => '<p>A wide range of human diseases result from haploinsufficiency, where the function of one of the two gene copies is lost. Here, we targeted the remaining functional copy of a haploinsufficient gene using CRISPR-mediated activation (CRISPRa) in and heterozygous mouse models to rescue their obesity phenotype. Transgenic-based CRISPRa targeting of the promoter or its distant hypothalamic enhancer up-regulated its expression from the endogenous functional allele in a tissue-specific manner, rescuing the obesity phenotype in heterozygous mice. To evaluate the therapeutic potential of CRISPRa, we injected CRISPRa-recombinant adeno-associated virus into the hypothalamus, which led to reversal of the obesity phenotype in and haploinsufficient mice. Our results suggest that endogenous gene up-regulation could be a potential strategy to treat altered gene dosage diseases.</p>',
'date' => '2019-01-18',
'pmid' => 'http://www.pubmed.gov/30545847',
'doi' => '10.1126/science.aau0629',
'modified' => '2019-06-07 09:05:53',
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×
