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  3. Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination

Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination

  • STAR Protoc. 2021 May 20;2(2):100548. doi: 10.1016/j.xpro.2021.100548.
Zhongshu Zhou 1 2,  Lin Ma 1 2 3 4,  Xiaoqing Zhang 1 5 2 6 3 4
Affiliations

Affiliations

  • 1 Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China.
  • 2 Key Laboratory of Neuroregeneration of Shanghai Universities, Tongji University School of Medicine, Shanghai 200092, China.
  • 3 Shanghai Institute of Stem Cell Research and Clinical Translation, Shanghai 200120, China.
  • 4 Brain and Spinal Cord Clinical Research Center, Tongji University School of Medicine, Shanghai 200092, China.
  • 5 Key Laboratory of Reconstruction and Regeneration of Spine and Spinal Cord Injury, Ministry of Education, Shanghai 200065, China.
  • 6 Tsingtao Advanced Research Institute, Tongji University, Qingdao 266071, China.
PMID: 34095862 DOI: 10.1016/j.xpro.2021.100548
Abstract

PAX6 is a key determinant of human neuroectoderm cell fate. Here, we describe a protocol for genome-scale CRISPR screening for use in genetically engineered human pluripotent stem cells (hPSCs). Using the germ layer reporter PAX6 and an inducible CRISPR/Cas9 knockout system, we describe how to identify lineage-specific preventing genes. This protocol can be applied for use with Other reporter genes to study cell fate determination in hPSCs. For complete details on the use and execution of this protocol, please refer to Xu et al. (2021).

Keywords

CRISPR; Cell Differentiation; High Throughput Screening; Stem Cells.

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