Modular vector assembly enables rapid assessment of emerging CRISPR technologies.

IF 11.1 Q1 CELL BIOLOGY
Abby V McGee, Yanjing V Liu, Audrey L Griffith, Zsofia M Szegletes, Bronte Wen, Carolyn Kraus, Nathan W Miller, Ryan J Steger, Berta Escude Velasco, Justin A Bosch, Jonathan D Zirin, Raghuvir Viswanatha, Erik J Sontheimer, Amy Goodale, Matthew A Greene, Thomas M Green, John G Doench
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引用次数: 0

Abstract

The diversity of CRISPR systems, coupled with scientific ingenuity, has led to an explosion of applications; however, to test newly described innovations in their model systems, researchers typically embark on cumbersome, one-off cloning projects to generate custom reagents that are optimized for their biological questions. Here, we leverage Golden Gate cloning to create the Fragmid toolkit, a modular set of CRISPR cassettes and delivery technologies, along with a web portal, resulting in a combinatorial platform that enables scalable vector assembly within days. We further demonstrate that multiple CRISPR technologies can be assessed in parallel in a pooled screening format using this resource, enabling the rapid optimization of both novel technologies and cellular models. These results establish Fragmid as a robust system for the rapid design of CRISPR vectors, and we anticipate that this assembly approach will be broadly useful for systematic development, comparison, and dissemination of CRISPR technologies.

模块化载体组装可快速评估新兴的 CRISPR 技术。
CRISPR 系统的多样性加上科学上的独创性,导致了应用的爆炸性增长;然而,为了在其模型系统中测试新描述的创新,研究人员通常会开展繁琐的一次性克隆项目,以生成针对其生物学问题进行优化的定制试剂。在这里,我们利用 "金门 "克隆技术创建了 Fragmid 工具包,这是一套模块化的 CRISPR 盒和传递技术以及一个门户网站,从而形成了一个组合平台,可在数天内完成可扩展的载体组装。我们进一步证明,利用这一资源,可以以集合筛选的形式并行评估多种 CRISPR 技术,从而快速优化新型技术和细胞模型。这些结果使 Fragmid 成为快速设计 CRISPR 载体的强大系统,我们预计这种组装方法将广泛用于 CRISPR 技术的系统开发、比较和推广。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
7.10
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