Cas13d: A New Molecular Scissor for Transcriptome Engineering.
Cas13d: A New Molecular Scissor for Transcriptome Engineering.
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DOI：
10.3389/fcell.2022.866800
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发表时间：
2022
影响因子：
5.5
通讯作者：
中科院分区：
文献类型：
--
作者：
在细菌和古细菌物种中发现的规则间隔回文重复序列（CRISPR）及其相关的Cas内切核酸酶使科学家能够修改，利用和彻底改变这种工具，用于任何物种的遗传改变。特别是II型CRISPR-Cas9系统在过去几十年中已被广泛研究并用于植物和哺乳动物系统中的精确和有效的DNA操作。此外，V型CRISPR-Cas 12（Cpf 1）系统的发现为原核生物、植物和动物中的DNA操作提供了更大的灵活性和精确性。然而，已经做出了很多努力来采用和利用上述CRISPR工具进行RNA操作，但是Cas9和Cas 12切割DNA的能力涉及对基因的脱靶效应的滋扰，因此可能不会在所有RNA靶向应用中采用。因此，人们开始寻找能够精确检测和操纵靶向RNA的新的、多样化的Cas效应子，这导致了靶向2类VI型CRISPR-Cas 13系统的新型RNA的发现。CRISPR-Cas 13系统由单个RNA引导的Cas 13效应核酸酶组成，其以可编程的方式仅靶向单链RNA（ssRNA）而不改变DNA。Cas 13效应子家族包括四个亚型（a-d），并且每个亚型具有独特的一级序列差异，除了两个共有的高等真核生物和原核生物核苷酸结合结构域（HEPN），其包括RNA酶基序，即R-X4-6-H。这两个HEPN结构域单独负责高效地执行靶向RNA切割活性。此外，最近的研究表明，与Cas 13酶家族的其他Cas 13内切核酸酶相比，Cas 13 d在哺乳动物系统中切割靶向RNA时表现出更高的效率和特异性。除此之外，Cas 13 d在结构上和功能上都表现出了优于其他Cas 13变体的额外优势，这使其成为RNA工程和编辑的突出和最高级工具。因此，考虑到Cas 13 d相对于先前表征的Cas 13亚型的优势，在这篇综述中，我们涵盖了VI型CRISPR-Cas 13 d系统的结构和机制特性，概述了目前报道的Cas 13 d的各种应用，以及改进基于Cas 13 d的诊断和治疗工具的前景。
The discovery of Clustered Regularly Interspaced Palindromic Repeats (CRISPR) and its associated Cas endonucleases in bacterial and archaeal species allowed scientists to modify, utilized, and revolutionize this tool for genetic alterations in any species. Especially the type II CRISPR-Cas9 system has been extensively studied and utilized for precise and efficient DNA manipulation in plant and mammalian systems over the past few decades. Further, the discovery of the type V CRISPR-Cas12 (Cpf1) system provides more flexibility and precision in DNA manipulation in prokaryotes, plants, and animals. However, much effort has been made to employ and utilize the above CRISPR tools for RNA manipulation but the ability of Cas9 and Cas12 to cut DNA involves the nuisance of off-target effects on genes and thus may not be employed in all RNA-targeting applications. Therefore, the search for new and diverse Cas effectors which can precisely detect and manipulate the targeted RNA begins and this led to the discovery of a novel RNA targeting class 2, type VI CRISPR-Cas13 system. The CRISPR-Cas13 system consists of single RNA-guided Cas13 effector nucleases that solely target single-stranded RNA (ssRNA) in a programmable way without altering the DNA. The Cas13 effectors family comprises four subtypes (a-d) and each subtype has distinctive primary sequence divergence except the two consensuses Higher eukaryotes and prokaryotes nucleotide-binding domain (HEPN) that includes RNase motifs i.e. R-X4-6-H. These two HEPN domains are solely responsible for executing targetable RNA cleavage activity with high efficiency. Further, recent studies have shown that Cas13d exhibits higher efficiency and specificity in cleaving targeted RNA in the mammalian system compared to other Cas13 endonucleases of the Cas13 enzyme family. In addition to that, Cas13d has shown additional advantages over other Cas13 variants, structurally as well as functionally which makes it a prominent and superlative tool for RNA engineering and editing. Therefore considering the advantages of Cas13d over previously characterized Cas13 subtypes, in this review, we encompass the structural and mechanistic properties of type VI CRISPR-Cas13d systems, an overview of the current reported various applications of Cas13d, and the prospects to improve Cas13d based tools for diagnostic and therapeutic purposes.
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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
影响因子：
64.8
作者：
Anders, Carolin;Niewoehner, Ole;Duerst, Alessia;Jinek, Martin
通讯作者：
Jinek, Martin
影响因子：
4
作者：
Desterro, JMP;Keegan, LR;Carmo-Fonseca, M
通讯作者：
Carmo-Fonseca, M
通过反式编码的小 RNA 和宿主因子 RNase III 进行 CRISPR RNA 成熟。
影响因子：
64.8
作者：
Deltcheva, Elitza;Chylinski, Krzysztof;Sharma, Cynthia M.;Gonzales, Karine;Chao, Yanjie;Pirzada, Zaid A.;Eckert, Maria R.;Vogel, Joerg;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
64.8
作者：
Abudayyeh OO;Gootenberg JS;Essletzbichler P;Han S;Joung J;Belanto JJ;Verdine V;Cox DBT;Kellner MJ;Regev A;Lander ES;Voytas DF;Ting AY;Zhang F
通讯作者：
Zhang F