Gene Editing in Mouse Lin-Sca1+Kit+ and Double Negative（DN）Primary Cell

doi:10.13560/j.cnki.biotech.bull.1985.2017-0903

Abstract

Abstract: The immune system of mice（Mus musculus）contains a large number of cell groups with different functions. In order to study the development mechanism of mouse immune system,it is necessary to establish efficient genetic manipulation techniques. Firstly CD4^-CD8^- double negative thymocytes（isolated from Cas9 knock-in mice）was chosen for this study,then sgRNAs（single guide RNA）were designed to target GFP and mouse CD4 cell surface antigen genes,and murine stem cell virus was used as a vector to deliver sgRNA into the cell,and by which gene editing was realized. The successful knockout of GFP and CD4 were detected by flow cytometry analysis. Based on these results,using Lin^-Sca1⁺Kit⁺ cell（isolated from Cas9 knock-in mice）as study materials,sgRNAs were designed and constructed with the same method for gene editing. After gene editing,the LSK cells did not differentiate into downstream T cells,but their myeloid differentiation potential was not affected. All these results suggest that we succeed in establishing mouse DN and LSK primary cell gene-editing platform.

Key words: mouse, immunology, stem cell, gene editing, CRISPR/Cas9

WANG Yue-qiang, Avinash Bhandoola. Gene Editing in Mouse Lin^-Sca1⁺Kit⁺ and Double Negative（DN）Primary Cell[J]. Biotechnology Bulletin, 2018, 34(5): 64-70.

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Gene Editing in Mouse Lin^-Sca1⁺Kit⁺ and Double Negative（DN）Primary Cell

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