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Pooled CRISPR-activation screening coupled with single-cell RNA-seq in mouse embryonic stem cells

2021; Elsevier BV; Volume: 2; Issue: 2 Linguagem: Inglês

10.1016/j.xpro.2021.100426

2666-1667

Celia Alda-Catalinas, Mélanie Eckersley-Maslin, Wolf Reik,

Pluripotent Stem Cells Research

CRISPR/Cas9 screens are a powerful approach to identify key regulators of biological processes. By combining pooled CRISPR/Cas9 screening with single-cell RNA-sequencing readout, individual perturbations can be assessed in parallel both comprehensively and at scale. Importantly, this allows gene function and regulation to be interrogated at a cellular level in an unbiased manner. Here, we present a protocol to perform pooled CRISPR-activation screens in mouse embryonic stem cells using 10× Genomics scRNA-seq as a readout. For complete information on the generation and use of this protocol, please refer to Alda-Catalinas et al. (2020).