Computational design of anti-CRISPR proteins with improved inhibition potency

被引：17

作者：

Mathony, Jan ^{[1
,2
,3
]}

Harteveld, Zander ^{[4
,5
]}

Schmelas, Carolin ^{[6
,7
,8
]}

zu Belzen, Julius Upmeier ^{[1
,2
,3
,9
]}

Aschenbrenner, Sabine ^{[1
,2
,3
,10
]}

Sun, Wei ^{[11
,12
]}

Hoffmann, Mareike D. ^{[1
,10
]}

Stengl, Christina ^{[1
]}

Scheck, Andreas ^{[4
,5
]}

Georgeon, Sandrine ^{[4
,5
]}

Rosset, Stephane ^{[4
,5
]}

Wang, Yanli ^{[11
,12
]}

Grimm, Dirk ^{[6
,7
,8
,13
,14
]}

Eils, Roland ^{[2
,3
,9
]}

Correia, Bruno E. ^{[4
,5
]}

Niopek, Dominik ^{[1
,9
]}

机构：

[1] Heidelberg Univ, Synthet Biol Grp, BioQuant Ctr, Heidelberg, Germany

[2] Berlin Inst Hlth, Digital Hlth Ctr, Berlin, Germany

[3] Charite, Berlin, Germany

[4] Ecole Polytech Fed Lausanne, Inst Bioengn, Lausanne, Switzerland

[5] Swiss Inst Bioinformat, Lausanne, Switzerland

[6] Univ Hosp Heidelberg, Dept Infect Dis, Virol, Heidelberg, Germany

[7] Heidelberg Univ, BioQuant Ctr, Heidelberg, Germany

[8] Heidelberg Univ, Cluster Excellence CellNetworks, Heidelberg, Germany

[9] Univ Hosp Heidelberg, Hlth Data Sci Unit, Heidelberg, Germany

[10] German Canc Res Ctr, Dept Theoret Bioinformat, Heidelberg, Germany

[11] Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing, Peoples R China

[12] Chinese Acad Sci, Inst Biophys, Key Lab RNA Biol, Beijing, Peoples R China

[13] German Ctr Infect Res DZIF, Heidelberg, Germany

[14] German Ctr Cardiovasc Res DZHK Partner Site Heide, Heidelberg, Germany

来源：

NATURE CHEMICAL BIOLOGY | 2020年 / 16卷 / 07期

基金：

瑞士国家科学基金会; 欧洲研究理事会;

关键词：

GENOME; ALGORITHM;

D O I：

10.1038/s41589-020-0518-9

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：

Anti-CRISPR (Acr) proteins are powerful tools to control CRISPR-Cas technologies. However, the available Acr repertoire is limited to naturally occurring variants. Here, we applied structure-based design on AcrIIC1, a broad-spectrum CRISPR-Cas9 inhibitor, to improve its efficacy on different targets. We first show that inserting exogenous protein domains into a selected AcrIIC1 surface site dramatically enhances inhibition of Neisseria meningitidis (Nme)Cas9. Then, applying structure-guided design to the Cas9-binding surface, we converted AcrIIC1 into AcrIIC1X, a potent inhibitor of the Staphylococcus aureus (Sau)Cas9, an orthologue widely applied for in vivo genome editing. Finally, to demonstrate the utility of AcrIIC1X for genome engineering applications, we implemented a hepatocyte-specific SauCas9 ON-switch by placing AcrIIC1X expression under regulation of microRNA-122. Our work introduces designer Acrs as important biotechnological tools and provides an innovative strategy to safeguard CRISPR technologies.

引用

页码：725 / +

页数：12

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