title: One-step generation of conditional and reversible gene knockouts.
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.
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.|Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: Amanda
authors.initials: A
authors.lastname: Andersson-Rolf
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Roxana C
authors.initials: RC
authors.lastname: Mustata
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.|Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Merenda
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Jihoon
authors.initials: J
authors.lastname: Kim
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UK.
authors.forename: Sajith
authors.initials: S
authors.lastname: Perera
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UK.
authors.forename: Tiago
authors.initials: T
authors.lastname: Grego
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UK.
authors.forename: Katie
authors.initials: K
authors.lastname: Andrews
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Katie
authors.initials: K
authors.lastname: Tremble
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.|Department of Biochemistry, University of Cambridge, Cambridge, UK.
authors.forename: José C R
authors.initials: JC
authors.lastname: Silva
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Juergen
authors.initials: J
authors.lastname: Fink
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UK.
authors.forename: William C
authors.initials: WC
authors.lastname: Skarnes
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.|Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: Bon-Kyoung
authors.initials: BK
authors.lastname: Koo
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology*; D000072669:Gene Editing / Q000379:methods*; D055786:Gene Knockout Techniques / Q000379:methods*; D016678:Genome / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D051176:beta Catenin / Q000235:genetics*
chemical_list: C495267:CTNNB1 protein, mouse; D051176:beta Catenin