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Nat Methods. 2017 Mar;14(3):287-289. doi: 10.1038/nmeth.4156. Epub 2017 Jan 30.

One-step generation of conditional and reversible gene knockouts.

Author information

1
Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
2
Department of Genetics, University of Cambridge, Cambridge, UK.
3
Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UK.
4
Department of Biochemistry, University of Cambridge, Cambridge, UK.

Abstract

Loss-of-function studies are key for investigating gene function, and CRISPR technology has made genome editing widely accessible in model organisms and cells. However, conditional gene inactivation in diploid cells is still difficult to achieve. Here, we present CRISPR-FLIP, a strategy that provides an efficient, rapid and scalable method for biallelic conditional gene knockouts in diploid or aneuploid cells, such as pluripotent stem cells, 3D organoids and cell lines, by co-delivery of CRISPR-Cas9 and a universal conditional intronic cassette.

PMID:
28135257
PMCID:
PMC5777571
DOI:
10.1038/nmeth.4156
[Indexed for MEDLINE]
Free PMC Article

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