Fanzor is a eukaryotic programmable RNA-guided endonuclease.
Fanzor is a eukaryotic programmable RNA-guided endonuclease.
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DOI：
10.1038/s41586-023-06356-2
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发表时间：
2023-08
期刊：
影响因子：
64.8
通讯作者：
Zhang, Feng
中科院分区：
文献类型：
--
作者：
Saito, Makoto;Xu, Peiyu;Faure, Guilhem;Maguire, Samantha;Kannan, Soumya;Altae-Tran, Han;Vo, Sam;Desimone, AnAn;Macrae, Rhiannon K.;Zhang, Feng
RNA引导的系统利用引导RNA和靶核酸序列之间的互补性来识别遗传元件，在原核生物和真核生物的生物过程中具有中心作用。例如，原核CRISPR-Cas系统为细菌和古细菌提供针对外源遗传元件的适应性免疫。Cas效应子如Cas9和Cas 12执行向导RNA依赖性DNA切割。虽然已经研究了一些真核生物的RNA引导系统，包括RNA干扰和核糖体RNA修饰，但真核生物是否具有RNA引导的核酸内切酶仍不清楚。最近，报道了一类新的原核RNA引导系统（称为OMEGA）。OMEGA效应子TnpB是Cas 12的假定祖先，具有RNA引导的核酸内切酶活性。TnpB也可能是真核转座子编码的Fanzor（Fz）蛋白的祖先，这提高了真核生物也配备有CRISPR-Cas或OMEGA样可编程RNA引导核酸内切酶的可能性。在这里，我们报告Fz的生化特性，表明它是一种RNA引导的DNA内切酶。我们还表明，Fz可以重新编程为人类基因组工程的应用。最后，我们解决了Spizellomyces punctatus Fz在2.7 ℃的结构，使用低温电子显微镜，显示Fz，TnpB和Cas 12之间的核心区域的保护，尽管不同的同源RNA结构。我们的研究结果表明，Fz是一个真核OMEGA系统，表明RNA引导的核酸内切酶存在于生命的所有三个领域。Fanzor被证明是一种RNA引导的DNA内切核酸酶，表明这种内切核酸酶存在于生命的所有领域，并表明基因组工程应用的潜在新工具。
RNA-guided systems, which use complementarity between a guide RNA and target nucleic acid sequences for recognition of genetic elements, have a central role in biological processes in both prokaryotes and eukaryotes. For example, the prokaryotic CRISPR–Cas systems provide adaptive immunity for bacteria and archaea against foreign genetic elements. Cas effectors such as Cas9 and Cas12 perform guide-RNA-dependent DNA cleavage. Although a few eukaryotic RNA-guided systems have been studied, including RNA interference and ribosomal RNA modification, it remains unclear whether eukaryotes have RNA-guided endonucleases. Recently, a new class of prokaryotic RNA-guided systems (termed OMEGA) was reported. The OMEGA effector TnpB is the putative ancestor of Cas12 and has RNA-guided endonuclease activity. TnpB may also be the ancestor of the eukaryotic transposon-encoded Fanzor (Fz) proteins, raising the possibility that eukaryotes are also equipped with CRISPR–Cas or OMEGA-like programmable RNA-guided endonucleases. Here we report the biochemical characterization of Fz, showing that it is an RNA-guided DNA endonuclease. We also show that Fz can be reprogrammed for human genome engineering applications. Finally, we resolve the structure of Spizellomyces punctatus Fz at 2.7 Å using cryogenic electron microscopy, showing the conservation of core regions among Fz, TnpB and Cas12, despite diverse cognate RNA structures. Our results show that Fz is a eukaryotic OMEGA system, demonstrating that RNA-guided endonucleases are present in all three domains of life. Fanzor is shown to be an RNA-guided DNA endonuclease, demonstrating that such endonucleases are found in all domains of life and indicating a potential new tool for genome engineering applications.
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影响因子：
64.8
作者：
Hirano, Seiichi;Kappel, Kalli;Altae-Tran, Han;Faure, Guilhem;Wilkinson, Max E.;Kannan, Soumya;Demircioglu, F. Esra;Yan, Rui;Shiozaki, Momoko;Yu, Zhiheng;Makarova, Kira S.;Koonin, Eugene, V;Macrae, Rhiannon K.;Zhang, Feng
通讯作者：
Zhang, Feng
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
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
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
4.9
作者：
Bao, Weidong;Jurka, Jerzy
通讯作者：
Jurka, Jerzy