Crystal structure of Staphylococcus aureus Cas9

Nishimasu, Hiroshi; Cong, Le; Yan, Winston X.; Ran, F. Ann; Zetsche, Bernd; Li, Yinqing; Kurabayashi, Arisa; Ishitani, Ryuichiro; Zhang, Feng; Nureki, Osamu

doi:10.1016/j.cell.2015.08.007

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Crystal structure of Staphylococcus aureus Cas9

Aug 27 2015
By Nishimasu, Hiroshi ; Cong, Le ; Yan, Winston X. ; ...
http://dx.doi.org/10.1016/j.cell.2015.08.007
Source: Cell. 162(5):1113-1126.

[PDF-2.76 MB]

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Details:

Alternative Title:

Cell
Personal Author:

Nishimasu, Hiroshi ; Cong, Le ; Yan, Winston X. ; Ran, F. Ann ; Zetsche, Bernd ; ... More +

Nishimasu, Hiroshi ; Cong, Le ; Yan, Winston X. ; Ran, F. Ann ; Zetsche, Bernd ; Li, Yinqing ; Kurabayashi, Arisa ; Ishitani, Ryuichiro ; Zhang, Feng ; Nureki, Osamu Less -
Description:

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 Å 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.
Subjects:

[+]

Amino Acid Sequence Article Bacterial Proteins CRISPR-Cas Systems Crystallography, X-Ray DNA Genetic Engineering Models, Molecular Molecular Sequence Data Protein Structure, Tertiary RNA, Guide Sequence Alignment Staphylococcus Aureus Streptococcus Pyogenes
Source:

Cell. 162(5):1113-1126.
Pubmed ID:

26317473
Pubmed Central ID:

PMC4670267
Document Type:

Journal Article
Funding:

1DP1-MH100706/DP/NCCDPHP CDC HHS/United States ; DP1 MH100706/MH/NIMH NIH HHS/United States ; R01 MH110049/MH/NIMH NIH HHS/United States ; T32 GM007753/GM/NIGMS NIH HHS/United States ; T32 GM008313/GM/NIGMS NIH HHS/United States ; ... More +

1DP1-MH100706/DP/NCCDPHP CDC HHS/United States ; DP1 MH100706/MH/NIMH NIH HHS/United States ; R01 MH110049/MH/NIMH NIH HHS/United States ; T32 GM007753/GM/NIGMS NIH HHS/United States ; T32 GM008313/GM/NIGMS NIH HHS/United States ; T32GM007753/GM/NIGMS NIH HHS/United States Less -
Volume:

162
Issue:

5
Collection(s):

CDC Public Access
Main Document Checksum:

[+]

urn:sha256:8f0f53a810208ba2c34c675bd4b64a6394ec977c6aecb63fedb88af1ef9f5a3a
Download URL:

https://stacks.cdc.gov/view/cdc/36575/cdc_36575_DS1.pdf
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