Large-scale multiplexed mosaic CRISPR perturbation in the whole organism

Bo Liu; Zhengyu Jing; Xiaoming Zhang; Yuxin Chen; Shaoshuai Mao; Ravinder Kaundal; Yan Zou; Ge Wei; Ying Zang; Xinxin Wang; Wenyang Lin; Minghui Di; Yiwen Sun; Qin Chen; Yongqin Li; Jing Xia; Jianlong Sun; Chao-Po Lin; Xingxu Huang; Tian Chi

doi:10.1016/j.cell.2022.06.039

Large-scale multiplexed mosaic CRISPR perturbation in the whole organism

Cell. 2022 Aug 4;185(16):3008-3024.e16. doi: 10.1016/j.cell.2022.06.039. Epub 2022 Jul 22.

Authors

Bo Liu¹, Zhengyu Jing¹, Xiaoming Zhang¹, Yuxin Chen¹, Shaoshuai Mao¹, Ravinder Kaundal², Yan Zou¹, Ge Wei¹, Ying Zang¹, Xinxin Wang¹, Wenyang Lin¹, Minghui Di¹, Yiwen Sun¹, Qin Chen¹, Yongqin Li¹, Jing Xia¹, Jianlong Sun¹, Chao-Po Lin¹, Xingxu Huang¹, Tian Chi³

Affiliations

¹ Gene Editing Center, School of Life Sciences and Technology, ShanghaiTech University, Shanghai 201210, China.
² Department of Immunobiology, Yale University Medical School, New Haven, CT 06520, USA.
³ Gene Editing Center, School of Life Sciences and Technology, ShanghaiTech University, Shanghai 201210, China; Department of Immunobiology, Yale University Medical School, New Haven, CT 06520, USA. Electronic address: chitian@shanghaitech.edu.cn.

PMID: 35870449
DOI: 10.1016/j.cell.2022.06.039

Abstract

Here, we report inducible mosaic animal for perturbation (iMAP), a transgenic platform enabling in situ CRISPR targeting of at least 100 genes in parallel throughout the mouse body. iMAP combines Cre-loxP and CRISPR-Cas9 technologies and utilizes a germline-transmitted transgene carrying a large array of individually floxed, tandemly linked gRNA-coding units. Cre-mediated recombination triggers expression of all the gRNAs in the array but only one of them per cell, converting the mice to mosaic organisms suitable for phenotypic characterization and also for high-throughput derivation of conventional single-gene perturbation lines via breeding. Using gRNA representation as a readout, we mapped a miniature Perturb-Atlas cataloging the perturbations of 90 genes across 39 tissues, which yields rich insights into context-dependent gene functions and provides a glimpse of the potential of iMAP in genome decoding.

Keywords: CAR T; Perturb-seq; cell atlas; immunotherapy; in vivo CRISPR screen; mouse cell Perturb-Atlas.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems* / genetics
Gene Editing
Genome
Mice
RNA, Guide, CRISPR-Cas Systems* / genetics
RNA, Guide, CRISPR-Cas Systems* / metabolism
Transgenes

Substances

RNA, Guide, CRISPR-Cas Systems