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Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides | |
2020-02-26 | |
发表期刊 | GENOME BIOLOGY |
ISSN | 1474-760X |
卷号 | 21期号:1 |
发表状态 | 已发表 |
DOI | 10.1186/s13059-020-01956-x |
摘要 | Background CRISPR-Cas9 has been developed as a therapeutic agent for various infectious and genetic diseases. In many clinically relevant applications, constitutively active CRISPR-Cas9 is delivered into human cells without a temporal control system. Excessive and prolonged expression of CRISPR-Cas9 can lead to elevated off-target cleavage. The need for modulating CRISPR-Cas9 activity over time and dose has created the demand of developing CRISPR-Cas off switches. Protein and small molecule-based CRISPR-Cas inhibitors have been reported in previous studies. Results We report the discovery of Cas9-inhibiting peptides from inoviridae bacteriophages. These peptides, derived from the periplasmic domain of phage major coat protein G8P (G8P(PD)), can inhibit the in vitro activity of Streptococcus pyogenes Cas9 (SpCas9) proteins in an allosteric manner. Importantly, the inhibitory activity of G8P(PD) on SpCas9 is dependent on the order of guide RNA addition. Ectopic expression of full-length G8P (G8P(FL)) or G8P(PD) in human cells can inactivate the genome-editing activity of SpyCas9 with minimum alterations of the mutation patterns. Furthermore, unlike the anti-CRISPR protein AcrII4A that completely abolishes the cellular activity of CRISPR-Cas9, G8P co-transfection can reduce the off-target activity of co-transfected SpCas9 while retaining its on-target activity. Conclusion G8Ps discovered in the current study represent the first anti-CRISPR peptides that can allosterically inactivate CRISPR-Cas9. This finding may provide insights into developing next-generation CRISPR-Cas inhibitors for precision genome engineering. |
关键词 | CRISPR-Cas9 Inoviridae bacteriophage Major coat protein G8P Allosteric inhibition Off-target activity |
收录类别 | SCI ; SCIE |
资助项目 | ShanghaiTech University Startup Fund[2019F0301-000-01] |
WOS研究方向 | Biotechnology & Applied Microbiology ; Genetics & Heredity |
WOS类目 | Biotechnology & Applied Microbiology ; Genetics & Heredity |
WOS记录号 | WOS:000517486500001 |
出版者 | BMC |
WOS关键词 | ANTI-CRISPR ; READ ALIGNMENT ; GENOME ; RNA ; CAS9 ; DISCOVERY ; SPECIFICITY ; MECHANISM ; SEQUENCE ; ACRIIA4 |
原始文献类型 | Article |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/119043 |
专题 | 生命科学与技术学院_硕士生 免疫化学研究所_特聘教授组_抗体结构学实验室 免疫化学研究所_特聘教授组_抗体化学实验室 免疫化学研究所_特聘教授组_功能筛选实验室 免疫化学研究所_公共科研平台_分析化学平台 iHuman研究所_PI研究组_刘志杰组 生命科学与技术学院_博士生 免疫化学研究所_PI研究组_刘佳组 免疫化学研究所_PI研究组_杨贝组 |
共同第一作者 | Wang, Shao-jie |
通讯作者 | Ma, Peixiang; Liu, Jia |
作者单位 | 1.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai 201210, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, CAS Ctr Excellence Mol Cell Sci, Shanghai, Peoples R China 4.Univ Chinese Acad Sci, Beijing, Peoples R China 5.Zhejiang Univ, Coll Life Sci, Hangzhou 310058, Zhejiang, Peoples R China 6.Yantai Univ, Key Lab Mol Pharmacol & Drug Evaluat Yantai Univ, Collaborat Innovat Ctr Adv Drug Delivery Syst & B, Sch Pharm,Minist Educ, Yantai 264005, Shandong, Peoples R China |
第一作者单位 | 免疫化学研究所; 生命科学与技术学院 |
通讯作者单位 | 免疫化学研究所 |
第一作者的第一单位 | 免疫化学研究所 |
推荐引用方式 GB/T 7714 | Cui, Yan-ru,Wang, Shao-jie,Chen, Jun,et al. Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides[J]. GENOME BIOLOGY,2020,21(1). |
APA | Cui, Yan-ru.,Wang, Shao-jie.,Chen, Jun.,Li, Jie.,Chen, Wenzhang.,...&Liu, Jia.(2020).Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides.GENOME BIOLOGY,21(1). |
MLA | Cui, Yan-ru,et al."Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides".GENOME BIOLOGY 21.1(2020). |
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