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

ABSTRACT: 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).

SUBMITTER: Alda-Catalinas C

PROVIDER: S-EPMC8055612 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Alda-Catalinas Celia C Eckersley-Maslin Melanie A MA Reik Wolf W

STAR protocols 20210409 2

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× Genom ...[more]

PMID: 33899013

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