Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

Van Trung Chu; Graf Robin; Wirtz TristanWeber TimmFavret JeremyLi XunPetsch KerstinNgoc Tung TranSieweke Michael H.Berek ClaudiaKuehn RalfRajewsky Klaus

首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America. >Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

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Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

机译：Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

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摘要

Applying clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-mediated mutagenesis to primary mouse immune cells, we used high-fidelity single guide RNAs (sgRNAs) designed with an sgRNA design tool (CrispRGold) to target genes in primary B cells, T cells, and macrophages isolated from a Cas9 transgenic mouse line. Using this system, we achieved an average knockout efficiency of 80 in B cells. On this basis, we established a robust small-scale CRISPR-mediated screen in these cells and identified genes essential for B-cell activation and plasma cell differentiation. This screening system does not require deep sequencing and may serve as a precedent for the application of CRISPR/Cas9 to primary mouse cells.

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《Proceedings of the National Academy of Sciences of the United States of America.》 |2016年第44期|12514-12519|共6页
作者
Van Trung Chu; Graf Robin; Wirtz TristanWeber TimmFavret JeremyLi XunPetsch KerstinNgoc Tung TranSieweke Michael H.Berek ClaudiaKuehn RalfRajewsky Klaus;
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作者单位

Deutsch Rheuma Forschungszentrum, D-10117 Berlin, Germany;

Max Delbruck Ctr Mol Med, D-13125 Berlin, Germany;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
CRISPR/Cas9; sgRNA design; knockout efficiency; primary cells; B cells;

Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL/6 Cas9 transgenic mouse line

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