Crystal Structure of Staphylococcus aureus Cas9

被引：349

作者：

Nishimasu, Hiroshi ^{[1
,2
]}

Cong, Le ^{[3
,4
,5
,6
]}

Yan, Winston X. ^{[3
,4
,5
,6
,7
]}

Ran, F. Ann ^{[3
,8
]}

Zetsche, Bernd ^{[3
,4
,5
,6
]}

Li, Yinqing ^{[3
,4
,5
,6
]}

Kurabayashi, Arisa ^{[1
]}

Ishitani, Ryuichiro ^{[1
]}

Zhang, Feng ^{[3
,4
,5
,6
]}

Nureki, Osamu ^{[1
]}

机构：

[1] Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Bunkyo Ku, Tokyo 1130032, Japan

[2] JST, PRESTO, Bunkyo Ku, Tokyo 1130032, Japan

[3] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[4] MIT, McGovern Inst Brain Res, Cambridge, MA 02139 USA

[5] MIT, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA

[6] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[7] Harvard Univ, Harvard Mit Div Hlth Sci & Technol, Grad Program Biophys, Sch Med, Boston, MA 02115 USA

[8] Harvard Univ, Soc Fellows, Cambridge, MA 02138 USA

来源：

CELL | 2015年 / 162卷 / 05期

关键词：

RNA-GUIDED ENDONUCLEASE; TARGET DNA RECOGNITION; STREPTOCOCCUS-THERMOPHILUS; HOLLIDAY JUNCTION; IMMUNE-SYSTEM; DUAL-RNA; CRISPR; COMPLEX; TRANSCRIPTION; PROKARYOTES;

D O I：

10.1016/j.cell.2015.08.007

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The RNA-guided DNA endonuclease Cas9 cleaves double-stranded DNA targets with a protospacer adjacent motif (PAM) and complementarity to the guide RNA. Recently, we harnessed Staphylococcus aureus Cas9 (SaCas9), which is significantly smaller than Streptococcus pyogenes Cas9 (SpCas9), to facilitate efficient in vivo genome editing. Here, we report the crystal structures of SaCas9 in complex with a single guide RNA (sgRNA) and its double-stranded DNA targets, containing the 5'-TTGAAT-3' PAM and the 5'-TTGGGT-3' PAM, at 2.6 and 2.7 angstrom resolutions, respectively. The structures revealed the mechanism of the relaxed recognition of the 5'-NNGRRT-3' PAM by SaCas9. A structural comparison of SaCas9 with SpCas9 highlighted both structural conservation and divergence, explaining their distinct PAM specificities and orthologous sgRNA recognition. Finally, we applied the structural information about this minimal Cas9 to rationally design compact transcriptional activators and inducible nucleases, to further expand the CRISPR-Cas9 genome editing toolbox.

引用

页码：1113 / 1126

页数：14

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