Tagmentation Buffer (1x)

Notice (8): Undefined variable: solution_of_interest [APP/View/Products/view.ctp, line 755]
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<p id="Par1" class="p p-first-last"><em>Streptococcus pyogenes</em><span>&nbsp;</span>Cas9 (SpCas9) and derived enzymes are widely used as genome editors, but their promiscuous nuclease activity often induces undesired mutations and chromosomal rearrangements. Several strategies for mapping off-target effects have emerged, but they suffer from limited sensitivity. To increase the detection sensitivity, we develop an off-target assessment workflow that uses Duplex Sequencing. The strategy increases sensitivity by one order of magnitude, identifying previously unknown SpCas9&rsquo;s off-target mutations in the humanized<span>&nbsp;</span><em>PCSK9</em><span>&nbsp;</span>mouse model. To reduce off-target risks, we perform a bioinformatic search and identify a high-fidelity Cas9 variant of the II-B subfamily from<span>&nbsp;</span><em>Parasutterella secunda</em><span>&nbsp;</span>(PsCas9). PsCas9 shows improved specificity as compared to SpCas9 across multiple tested sites, both in vitro and in vivo, including the<span>&nbsp;</span><em>PCSK9</em><span>&nbsp;</span>site. In the future, while PsCas9 will offer an alternative to SpCas9 for research and clinical use, the Duplex Sequencing workflow will&nbsp;enable a more sensitive assessment of Cas9 editing outcomes.</p>
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<div class="sec"><strong class="kwd-title">Subject terms:<span>&nbsp;</span></strong><span class="kwd-text">Genetic engineering, Gene therapy, CRISPR-Cas9 genome editing</span></div>
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<p id="Par1" class="p p-first-last"><em>Streptococcus pyogenes</em><span>&nbsp;</span>Cas9 (SpCas9) and derived enzymes are widely used as genome editors, but their promiscuous nuclease activity often induces undesired mutations and chromosomal rearrangements. Several strategies for mapping off-target effects have emerged, but they suffer from limited sensitivity. To increase the detection sensitivity, we develop an off-target assessment workflow that uses Duplex Sequencing. The strategy increases sensitivity by one order of magnitude, identifying previously unknown SpCas9&rsquo;s off-target mutations in the humanized<span>&nbsp;</span><em>PCSK9</em><span>&nbsp;</span>mouse model. To reduce off-target risks, we perform a bioinformatic search and identify a high-fidelity Cas9 variant of the II-B subfamily from<span>&nbsp;</span><em>Parasutterella secunda</em><span>&nbsp;</span>(PsCas9). PsCas9 shows improved specificity as compared to SpCas9 across multiple tested sites, both in vitro and in vivo, including the<span>&nbsp;</span><em>PCSK9</em><span>&nbsp;</span>site. In the future, while PsCas9 will offer an alternative to SpCas9 for research and clinical use, the Duplex Sequencing workflow will&nbsp;enable a more sensitive assessment of Cas9 editing outcomes.</p>
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<div class="sec"><strong class="kwd-title">Subject terms:<span>&nbsp;</span></strong><span class="kwd-text">Genetic engineering, Gene therapy, CRISPR-Cas9 genome editing</span></div>
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View::render() - CORE/Cake/View/View.php, line 473
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Notice (8): Undefined variable: header [APP/View/Products/view.ctp, line 755]
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			'date' => '2024-11-07',
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</div>
<div class="sec"><strong class="kwd-title">Subject terms:<span>&nbsp;</span></strong><span class="kwd-text">Genetic engineering, Gene therapy, CRISPR-Cas9 genome editing</span></div>
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</div>
<div class="sec"><strong class="kwd-title">Subject terms:<span>&nbsp;</span></strong><span class="kwd-text">Genetic engineering, Gene therapy, CRISPR-Cas9 genome editing</span></div>
</div>
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View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
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Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
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Notice (8): Undefined variable: message [APP/View/Products/view.ctp, line 755]
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<div class="sec"><strong class="kwd-title">Subject terms:<span>&nbsp;</span></strong><span class="kwd-text">Genetic engineering, Gene therapy, CRISPR-Cas9 genome editing</span></div>
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118

Notice (8): Undefined variable: campaign_id [APP/View/Products/view.ctp, line 755]
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	'name' => 'Tagmentation Buffer (1x) SDS ES es',
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	'id' => '4879',
	'name' => 'A Type II-B Cas9 nuclease with minimized off-targets and reduced chromosomal translocations in vivo',
	'authors' => 'Bestas B. et al.',
	'description' => '<div id="Abs1" lang="en" class="tsec sec">
<div>
<p id="Par1" class="p p-first-last"><em>Streptococcus pyogenes</em><span>&nbsp;</span>Cas9 (SpCas9) and derived enzymes are widely used as genome editors, but their promiscuous nuclease activity often induces undesired mutations and chromosomal rearrangements. Several strategies for mapping off-target effects have emerged, but they suffer from limited sensitivity. To increase the detection sensitivity, we develop an off-target assessment workflow that uses Duplex Sequencing. The strategy increases sensitivity by one order of magnitude, identifying previously unknown SpCas9&rsquo;s off-target mutations in the humanized<span>&nbsp;</span><em>PCSK9</em><span>&nbsp;</span>mouse model. To reduce off-target risks, we perform a bioinformatic search and identify a high-fidelity Cas9 variant of the II-B subfamily from<span>&nbsp;</span><em>Parasutterella secunda</em><span>&nbsp;</span>(PsCas9). PsCas9 shows improved specificity as compared to SpCas9 across multiple tested sites, both in vitro and in vivo, including the<span>&nbsp;</span><em>PCSK9</em><span>&nbsp;</span>site. In the future, while PsCas9 will offer an alternative to SpCas9 for research and clinical use, the Duplex Sequencing workflow will&nbsp;enable a more sensitive assessment of Cas9 editing outcomes.</p>
</div>
<div class="sec"><strong class="kwd-title">Subject terms:<span>&nbsp;</span></strong><span class="kwd-text">Genetic engineering, Gene therapy, CRISPR-Cas9 genome editing</span></div>
</div>
<div id="Abs2" lang="en" class="tsec sec"></div>',
	'date' => '2023-09-06',
	'pmid' => 'https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10482872/',
	'doi' => '10.1038/s41467-023-41240-7',
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$externalLink = ' <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10482872/" target="_blank"><i class="fa fa-external-link"></i></a>'
include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118