2. Academic Validation
  3. Large-scale multiplexed mosaic CRISPR perturbation in the whole organism

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.
Bo Liu 1,  Zhengyu Jing 1,  Xiaoming Zhang 1,  Yuxin Chen 1,  Shaoshuai Mao 1,  Ravinder Kaundal 2,  Yan Zou 1,  Ge Wei 1,  Ying Zang 1,  Xinxin Wang 1,  Wenyang Lin 1,  Minghui Di 1,  Yiwen Sun 1,  Qin Chen 1,  Yongqin Li 1,  Jing Xia 1,  Jianlong Sun 1,  Chao-Po Lin 1,  Xingxu Huang 1,  Tian Chi 3
Affiliations

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.

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