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Pooled CRISPR screening with single-cell transcriptome readout.

ABSTRACT: CRISPR-based genetic screens are accelerating biological discovery, but current methods have inherent limitations. Widely used pooled screens are restricted to simple readouts including cell proliferation and sortable marker proteins. Arrayed screens allow for comprehensive molecular readouts such as transcriptome profiling, but at much lower throughput. Here we combine pooled CRISPR screening with single-cell RNA sequencing into a broadly applicable workflow, directly linking guide RNA expression to transcriptome responses in thousands of individual cells. Our method for CRISPR droplet sequencing (CROP-seq) enables pooled CRISPR screens with single-cell transcriptome resolution, which will facilitate high-throughput functional dissection of complex regulatory mechanisms and heterogeneous cell populations.

SUBMITTER: Datlinger P

PROVIDER: S-EPMC5334791 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Pooled CRISPR screening with single-cell transcriptome readout.

Datlinger Paul P Rendeiro André F AF Schmidl Christian C Krausgruber Thomas T Traxler Peter P Klughammer Johanna J Schuster Linda C LC Kuchler Amelie A Alpar Donat D Bock Christoph C

Nature methods 20170118 3

CRISPR-based genetic screens are accelerating biological discovery, but current methods have inherent limitations. Widely used pooled screens are restricted to simple readouts including cell proliferation and sortable marker proteins. Arrayed screens allow for comprehensive molecular readouts such as transcriptome profiling, but at much lower throughput. Here we combine pooled CRISPR screening with single-cell RNA sequencing into a broadly applicable workflow, directly linking guide RNA expressi ...[more]

PMID: 28099430

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