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Pooled CRISPR screening with single-cell transcriptome read-out

2016; Cold Spring Harbor Laboratory; Linguagem: Inglês

10.1101/083774

Paul Datlinger, Christian Schmidl, André F. Rendeiro, Peter Traxler, Johanna Klughammer, Linda C. Schuster, Christoph Bock,

RNA and protein synthesis mechanisms

Abstract CRISPR-based genetic screens have revolutionized the search for new gene functions and biological mechanisms. However, widely used pooled screens are limited to simple read-outs of cell proliferation or the production of a selectable marker protein. Arrayed screens allow for more complex molecular read-outs such as transcriptome profiling, but they provide much lower throughput. Here we demonstrate CRISPR genome editing together with single-cell RNA sequencing as a new screening paradigm that combines key advantages of pooled and arrayed screens. This approach allowed us to link guide-RNA expression to the associated transcriptome responses in thousands of single cells using a straightforward and broadly applicable screening workflow.