Quantification of Genome Editing and Transcriptional Control Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in Human Cells.
Quantification of Genome Editing and Transcriptional Control Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in Human Cells.
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DOI：
10.1021/acssynbio.2c00156
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发表时间：
2022-10-21
影响因子：
4.7
通讯作者：
Hilton, Isaac B.
中科院分区：
文献类型：
--
作者：
Escobar, Mario;Li, Jing;Patel, Aditi;Liu, Shizhe;Xu, Qi;Hilton, Isaac B.
CRISPR/Cas技术彻底改变了重新设计基因组信息和定制内源基因表达的能力。然而，新的CRISPR/Cas系统的发现和开发导致这些技术在人类细胞中的相对功效缺乏明确性。这种缺陷使得CRISPR/Cas技术在人类细胞中的最佳选择具有不必要的挑战性，这反过来又阻碍了它们的采用，从而最终限制了它们的实用性。在这里，我们设计了一系列内源性测试平台系统，以系统地量化和比较跨越II型和V型CRISPR/Cas家族的10种不同天然和工程化Cas蛋白变体的基因组编辑，CRISPRi和CRISPRa能力。我们表明，尽管所有Cas蛋白变体都能够在人类细胞中进行基因组编辑和转录控制，但存在层次结构，特别是对于基因组编辑和CRISPRa应用，其中Cas9 ≥ Cas 12 a> Cas 12 e/Cas 12 j。我们的研究结果还强调了我们的模块化测试平台的实用性，可以快速系统地量化人类细胞中几乎任何天然或工程基因组靶向Cas蛋白的功能。
CRISPR/Cas technologies have revolutionized the ability to redesign genomic information and tailor endogenous gene expression. Nevertheless, the discovery and development of new CRISPR/Cas systems has resulted in a lack of clarity surrounding the relative efficacies among these technologies in human cells. This deficit makes the optimal selection of CRISPR/Cas technologies in human cells unnecessarily challenging, which in turn hampers their adoption, and thus ultimately limits their utility. Here, we designed a series of endogenous testbed systems to methodically quantify and compare the genome editing, CRISPRi, and CRISPRa capabilities among 10 different natural and engineered Cas protein variants spanning Type II and Type V CRISPR/Cas families. We show that although all Cas protein variants are capable of genome editing and transcriptional control in human cells, hierarchies exist, particularly for genome editing and CRISPRa applications, wherein Cas9 ≥ Cas12a > Cas12e/Cas12j. Our findings also highlight the utility of our modular testbed platforms to rapidly and systematically quantify the functionality of practically any natural or engineered genomic-targeting Cas protein in human cells.
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影响因子：
64.8
作者：
Konermann S;Brigham MD;Trevino AE;Joung J;Abudayyeh OO;Barcena C;Hsu PD;Habib N;Gootenberg JS;Nishimasu H;Nureki O;Zhang F
通讯作者：
Zhang F
影响因子：
46.9
作者：
Kleinstiver BP;Tsai SQ;Prew MS;Nguyen NT;Welch MM;Lopez JM;McCaw ZR;Aryee MJ;Joung JK
通讯作者：
Joung JK
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
14.9
作者：
Brinkman EK;Chen T;Amendola M;van Steensel B
通讯作者：
van Steensel B
影响因子：
46.9
作者：
Kim DY;Lee JM;Moon SB;Chin HJ;Park S;Lim Y;Kim D;Koo T;Ko JH;Kim YS
通讯作者：
Kim YS