Discovery of Diverse CRISPR-Cas Systems and Expansion of the Genome Engineering Toolbox.
Discovery of Diverse CRISPR-Cas Systems and Expansion of the Genome Engineering Toolbox.
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CRISPR-CAS系统多样性的发现和基因组工程工具箱的扩展
DOI：
10.1021/acs.biochem.3c00159
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发表时间：
2023-12-19
期刊：
影响因子：
2.9
通讯作者：
Abudayyeh, Omar O.
中科院分区：
文献类型：
--
作者：
Koonin, Eugene V.;Gootenberg, Jonathan S.;Abudayyeh, Omar O.
CRISPR系统通过不同的效应机制介导细菌和古细菌中的适应性免疫，并且由于其与RNA向导的轻松重编程，已被重新用于治疗和诊断中的多功能应用。RNA引导的CRISPR-Cas靶向和干扰由效应子介导，所述效应子是1类系统中的多亚基复合物的组分或2类中的多结构域单效应蛋白。紧凑型2类CRISPR系统已被广泛用于多种应用，特别是基因组编辑，导致分子生物学和生物技术工具包的转变。最初仅限于Cas9核酸酶的2类效应酶的多样性通过计算基因组和宏基因组挖掘而大幅扩展，以包括Cas 12和Cas 13的许多变体，为开发通用的正交分子工具提供了底物。这些不同CRISPR效应子的表征揭示了许多新特征，包括扩展靶向空间的独特前间区序列相邻基序（PAM）、改进的编辑特异性、RNA而不是DNA靶向、较小的crRNA、交错和钝端切割、微型酶、混杂的RNA和DNA切割等。例如利用VI型效应物Cas 13的混杂RNA酶活性进行超灵敏的核酸检测。1类CRISPR系统也已被用于基因组编辑，尽管在表达和递送多蛋白1类效应物方面存在挑战。CRISPR酶的丰富多样性导致了基因组编辑工具箱的快速成熟，具有基因敲除、碱基编辑、引物编辑、基因插入、DNA成像、表观遗传调节、转录调节和RNA编辑等功能。结合效应蛋白和相关RNA的合理设计和工程改造，CRISPR和相关细菌RNA引导系统的天然多样性为扩大分子生物学和生物技术的工具库提供了巨大的资源。
CRISPR systems mediate adaptive immunity in bacteria and archaea through diverse effector mechanisms and have been repurposed for versatile applications in therapeutics and diagnostics thanks to their facile reprogramming with RNA guides. RNA-guided CRISPR-Cas targeting and interference are mediated by effectors that are either components of multisubunit complexes in class 1 systems or multidomain single-effector proteins in class 2. The compact class 2 CRISPR systems have been broadly adopted for multiple applications, especially genome editing, leading to a transformation of the molecular biology and biotechnology toolkit. The diversity of class 2 effector enzymes, initially limited to the Cas9 nuclease, was substantially expanded via computational genome and metagenome mining to include numerous variants of Cas12 and Cas13, providing substrates for the development of versatile, orthogonal molecular tools. Characterization of these diverse CRISPR effectors uncovered many new features, including distinct protospacer adjacent motifs (PAMs) that expand the targeting space, improved editing specificity, RNA rather than DNA targeting, smaller crRNAs, staggered and blunt end cuts, miniature enzymes, promiscuous RNA and DNA cleavage, etc. These unique properties enabled multiple applications, such as harnessing the promiscuous RNase activity of the type VI effector, Cas13, for supersensitive nucleic acid detection. class 1 CRISPR systems have been adopted for genome editing, as well, despite the challenge of expressing and delivering the multiprotein class 1 effectors. The rich diversity of CRISPR enzymes led to rapid maturation of the genome editing toolbox, with capabilities such as gene knockout, base editing, prime editing, gene insertion, DNA imaging, epigenetic modulation, transcriptional modulation, and RNA editing. Combined with rational design and engineering of the effector proteins and associated RNAs, the natural diversity of CRISPR and related bacterial RNA-guided systems provides a vast resource for expanding the repertoire of tools for molecular biology and biotechnology.
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影响因子：
64.8
作者：
Anders, Carolin;Niewoehner, Ole;Duerst, Alessia;Jinek, Martin
通讯作者：
Jinek, Martin
DOI：
10.1126/science.aaf5573
发表时间：
2016-08-05
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Abudayyeh OO;Gootenberg JS;Konermann S;Joung J;Slaymaker IM;Cox DB;Shmakov S;Makarova KS;Semenova E;Minakhin L;Severinov K;Regev A;Lander ES;Koonin EV;Zhang F
通讯作者：
Zhang F
DOI：
10.1016/j.omtm.2020.09.001
发表时间：
2020-12-11
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
Backstrom JR;Sheng J;Wang MC;Bernardo-Colón A;Rex TS
通讯作者：
Rex TS
影响因子：
64.8
作者：
Ackerman, Cheri M.;Myhrvold, Cameron;Sabeti, Pardis C.
通讯作者：
Sabeti, Pardis C.
影响因子：
28.3
作者：
Barrangou, Rodolphe;Horvath, Philippe
通讯作者：
Horvath, Philippe