Genome editing with type II-C CRISPR-Cas9 systems from Neisseria meningitidis in rice.
Genome editing with type II-C CRISPR-Cas9 systems from Neisseria meningitidis in rice.
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DOI：
10.1111/pbi.13716
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发表时间：
2022-03
影响因子：
13.8
通讯作者：
Wei P
中科院分区：
文献类型：
--
作者：
Xu R;Qin R;Xie H;Li J;Liu X;Zhu M;Sun Y;Yu Y;Lu P;Wei P
最近从脑膜炎奈瑟菌中鉴定出两种II-C型Cas9直系同源物（Nm 1Cas 9和Nm 2Cas 9），并已广泛用于哺乳动物细胞，但这些NmCas 9直系同源物或其他II-C型Cas9蛋白是否可以介导植物中的基因组编辑仍不清楚。在这项研究中，我们开发并优化了NmCas 9 s的植物靶向诱变系统。分别用Nm 1Cas 9和Nm 2Cas 9实现了在具有N4 GATT和N4 CC原型间隔区邻近基序（PAM）的靶标处的有效基因组编辑。这些结果表明，可以从具有不同PAM偏好的II-C型Cas9开发出高度活性的编辑系统，从而为扩大植物基因组编辑的范围提供了可靠的策略。碱基编辑器（BE）从NmCas 9进一步开发。TadA*-7.10的腺嘌呤BE（ABE）和大鼠APOBEC 1（rAPO 1）或人APOBEC 3a（hA 3A）的胞嘧啶BE（CBE）的编辑效率非常有限，而TadA-8 e的ABE和海岩巨虫胞苷脱氨酶1（PmCDA 1）的CBE在相同靶点上表现出显著改善的性能。此外，我们发现来自人Rad 51蛋白的单链DNA结合结构域的融合增强了NmCas 9的rAPO 1-CBE的碱基编辑能力。总之，我们的研究结果表明，NmCas 9 s或其他II-C型Cas9 s的工程改造可以为作物基因组编辑提供有用的替代方案。
Two type II‐C Cas9 orthologs (Nm1Cas9 and Nm2Cas9) were recently identified from Neisseria meningitidis and have been extensively used in mammalian cells, but whether these NmCas9 orthologs or other type II‐C Cas9 proteins can mediate genome editing in plants remains unclear. In this study, we developed and optimized targeted mutagenesis systems from NmCas9s for plants. Efficient genome editing at the target with N4GATT and N4CC protospacer adjacent motifs (PAMs) was achieved with Nm1Cas9 and Nm2Cas9 respectively. These results indicated that a highly active editing system could be developed from type II‐C Cas9s with distinct PAM preferences, thus providing a reliable strategy to extend the scope of genome editing in plants. Base editors (BEs) were further developed from the NmCas9s. The editing efficiency of adenine BEs (ABEs) of TadA*‐7.10 and cytosine BEs (CBEs) of rat APOBEC1 (rAPO1) or human APOBEC3a (hA3A) were extremely limited, whereas ABEs of TadA‐8e and CBEs of Petromyzon marinus cytidine deaminase 1 (PmCDA1) exhibited markedly improved performance on the same targets. In addition, we found that fusion of a single‐stranded DNA‐binding domain from the human Rad51 protein enhanced the base editing capability of rAPO1‐CBEs of NmCas9s. Together, our results suggest that the engineering of NmCas9s or other type II‐C Cas9s can provide useful alternatives for crop genome editing.
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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：
Fedorova, Iana;Arseniev, Anatolii;Severinov, Konstantin
通讯作者：
Severinov, Konstantin
DOI：
10.1126/science.abb1390
发表时间：
2020-07-31
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Lapinaite A;Knott GJ;Palumbo CM;Lin-Shiao E;Richter MF;Zhao KT;Beal PA;Liu DR;Doudna JA
通讯作者：
Doudna JA
影响因子：
4.6
作者：
Kaya H;Mikami M;Endo A;Endo M;Toki S
通讯作者：
Toki S
影响因子：
12.3
作者：
Li C;Zong Y;Wang Y;Jin S;Zhang D;Song Q;Zhang R;Gao C
通讯作者：
Gao C