用于在工程谱系报告 hPSC 中进行全基因组规模的 CRISPR 筛选以研究细胞命运决定的方案。
Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination.
机构信息
Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China.
Key Laboratory of Neuroregeneration of Shanghai Universities, Tongji University School of Medicine, Shanghai 200092, China.
出版信息
STAR Protoc. 2021 May 20;2(2):100548. doi: 10.1016/j.xpro.2021.100548. eCollection 2021 Jun 18.
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 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).
是人类神经外胚层细胞命运的关键决定因素。在这里，我们描述了一种用于基因工程人类多能干细胞（hPSC）的全基因组 CRISPR 筛选方案。使用胚层报告基因和诱导型 CRISPR/Cas9 敲除系统，我们描述了如何鉴定谱系特异性预防基因。该方案可用于其他报告基因，以研究 hPSC 中的细胞命运决定。有关此方案的使用和执行的完整详细信息，请参阅 Xu 等人。（2021 年）。
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