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CRISPR-表面组:一种用于设计靶向细胞表面蛋白的高效sgRNA的在线工具。

CRISPR-surfaceome: An online tool for designing highly efficient sgRNAs targeting cell surface proteins.

作者信息

Mei Hong, Gu Qian, Wang Wei, Meng Yu, Jiang Lichun, Liu Jia

机构信息

Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai 201210, China.

School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.

出版信息

Comput Struct Biotechnol J. 2022 Jul 18;20:3833-3838. doi: 10.1016/j.csbj.2022.07.026. eCollection 2022.

Abstract

CRISPR-based genome-editing tools have emerged as an efficient tool for functional genomics studies. Online tools and databases have been developed to facilitate the design and selection of CRISPR single guide RNA (sgRNA) for gene modifications. However, to the best of our knowledge, none of these tools or database are designated to cell surface proteins. In a previous study, we described the development and application of surfaceome CRISPR libraries targeting to cell surface proteins on human cells. Here, we present the design and construction of an online tool and database (https://crispr-surfaceome.siais.shanghaitech.edu.cn/home), named CRISPR-Surfaceome, for the design of highly efficient sgRNA targeting to the surface proteins on human cells. To show case and validate the efficiencies of sgRNAs designed by this online tool, we chose gene for knockout studies and found that all the 10 designed sgRNAs could efficiently generate knockout cells, with more than 80% gene disruption rates. These knockout cells were found to be resistant to the infection of rhinovirus (RV), which utilizes ICAM-1 as the receptor. Therefore, CRISPR-Surfaceome can serve the research community for the functional genomics studies on cell surface proteins, such as identification of pathogen receptors and discovery of drug targets.

摘要

基于CRISPR的基因组编辑工具已成为功能基因组学研究的有效工具。人们开发了在线工具和数据库,以促进用于基因修饰的CRISPR单向导RNA(sgRNA)的设计和选择。然而,据我们所知,这些工具或数据库都未针对细胞表面蛋白。在先前的一项研究中,我们描述了针对人类细胞表面蛋白的表面组CRISPR文库的开发和应用。在此,我们展示了一个名为CRISPR-Surfaceome的在线工具和数据库(https://crispr-surfaceome.siais.shanghaitech.edu.cn/home)的设计和构建,该工具用于设计针对人类细胞表面蛋白的高效sgRNA。为了展示和验证该在线工具设计的sgRNA的效率,我们选择了用于敲除研究的基因,发现所有设计的10个sgRNA都能有效地产生敲除细胞,基因破坏率超过80%。这些敲除细胞被发现对利用ICAM-1作为受体的鼻病毒(RV)感染具有抗性。因此,CRISPR-Surfaceome可为研究界提供服务,用于细胞表面蛋白的功能基因组学研究,如病原体受体的鉴定和药物靶点的发现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05c7/9307495/c0ed54801c9a/ga1.jpg

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