Programmable RNA-guided DNA endonucleases are widespread in eukaryotes and their viruses.
Programmable RNA-guided DNA endonucleases are widespread in eukaryotes and their viruses.
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DOI：
10.1126/sciadv.adk0171
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发表时间：
2023-09-29
期刊：
影响因子：
13.6
通讯作者：
Abudayyeh, Omar O.
中科院分区：
文献类型：
--
作者：
Jiang, Kaiyi;Lim, Justin;Sgrizzi, Samantha;Trinh, Michael;Kayabolen, Alisan;Yutin, Natalya;Bao, Weidong;Kato, Kazuki;Koonin, Eugene V.;Gootenberg, Jonathan S.;Abudayyeh, Omar O.
可编程rna引导的DNA核酸酶在原核生物中发挥着许多作用，但它们在原核生物外的传播程度尚不清楚。Fanzors是原核TnpB蛋白的真核同源物，已在真核生物和大型病毒的基因组中检测到，但其在真核生物中的活性和功能尚不清楚。在这里，我们将Fanzors描述为rna可编程DNA内切酶，使用生化和细胞证据。我们发现不同的Fanzors经常与各种真核转座酶相关联。Fanzors的进化重建揭示了真核生物中含有ruvc的TnpB同源物的多重辐射。Fanzor基因捕获内含子和蛋白质获得核定位信号，表明在真核细胞中广泛、长期地适应功能。Fanzor核酸酶含有RuvC结构域的重排催化位点，类似于TnpBs的一个独特子集，缺乏侧支裂解活性。我们证明了Fanzors可以用于人类细胞的基因组编辑，突出了这些广泛存在的真核rna引导核酸酶在生物技术应用中的潜力。Fanzor是一种广泛存在于真核生物及其病毒中的rna引导DNA内切酶。
Programmable RNA-guided DNA nucleases perform numerous roles in prokaryotes, but the extent of their spread outside prokaryotes is unclear. Fanzors, the eukaryotic homolog of prokaryotic TnpB proteins, have been detected in genomes of eukaryotes and large viruses, but their activity and functions in eukaryotes remain unknown. Here, we characterize Fanzors as RNA-programmable DNA endonucleases, using biochemical and cellular evidence. We found diverse Fanzors that frequently associate with various eukaryotic transposases. Reconstruction of Fanzors evolution revealed multiple radiations of RuvC-containing TnpB homologs in eukaryotes. Fanzor genes captured introns and proteins acquired nuclear localization signals, indicating extensive, long-term adaptation to functioning in eukaryotic cells. Fanzor nucleases contain a rearranged catalytic site of the RuvC domain, similar to a distinct subset of TnpBs, and lack collateral cleavage activity. We demonstrate that Fanzors can be harnessed for genome editing in human cells, highlighting the potential of these widespread eukaryotic RNA-guided nucleases for biotechnology applications. Fanzor is an RNA-guided DNA endonuclease widespread in eukaryotes and their viruses.
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DOI：
10.1126/science.aav4294
发表时间：
2018-11-16
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Harrington LB;Burstein D;Chen JS;Paez-Espino D;Ma E;Witte IP;Cofsky JC;Kyrpides NC;Banfield JF;Doudna JA
通讯作者：
Doudna JA
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：
4.9
作者：
Bao W;Kojima KK;Kohany O
通讯作者：
Kohany O
影响因子：
64.8
作者：
Saito, Makoto;Xu, Peiyu;Faure, Guilhem;Maguire, Samantha;Kannan, Soumya;Altae-Tran, Han;Vo, Sam;Desimone, AnAn;Macrae, Rhiannon K.;Zhang, Feng
通讯作者：
Zhang, Feng
影响因子：
64.8
作者：
Nakagawa, Ryoya;Hirano, Hisato;Omura, Satoshi N.;Nety, Suchita;Kannan, Soumya;Altae-Tran, Han;Yao, Xiao;Sakaguchi, Yuriko;Ohira, Takayuki;Wu, Wen Y.;Nakayama, Hiroshi;Shuto, Yutaro;Tanaka, Tatsuki;Sano, Fumiya K.;Kusakizako, Tsukasa;Kise, Yoshiaki;Itoh, Yuzuru;Dohmae, Naoshi;van der Oost, John;Suzuki, Tsutomu;Zhang, Feng;Nureki, Osamu
通讯作者：
Nureki, Osamu