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Journal Article
Pooled CRISPR screening with single-cell transcriptome readout
2017
Overview
CROP-seq enables pooled CRISPR screens for complex transcriptome signatures by making gRNA expression detectable in single-cell RNA sequencing.
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.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ 38
/ 38/39
/ 38/44
/ 38/47
/ 42/40
/ 42/41
/ 42/70
/ 45/77
/ 45/91
/ Biomedical Engineering/Biotechnology
/ Clustered Regularly Interspaced Short Palindromic Repeats - genetics
/ Gene Expression Profiling - methods
/ Genetics
/ High-Throughput Nucleotide Sequencing - methods
/ Humans
/ Identification and classification
/ RNA
/ RNA, Guide, CRISPR-Cas Systems - genetics
/ Sequence Analysis, RNA - methods
