title: Inducible in vivo genome editing with CRISPR-Cas9.
abstract: CRISPR-Cas9-based genome editing enables the rapid genetic manipulation of any genomic locus without the need for gene targeting by homologous recombination. Here we describe a conditional transgenic approach that allows temporal control of CRISPR-Cas9 activity for inducible genome editing in adult mice. We show that doxycycline-regulated Cas9 induction enables widespread gene disruption in multiple tissues and that limiting the duration of Cas9 expression or using a Cas9(D10A) (Cas9n) variant can regulate the frequency and size of target gene modifications, respectively. Further, we show that this inducible CRISPR (iCRISPR) system can be used effectively to create biallelic mutation in multiple target loci and, thus, provides a flexible and fast platform to study loss-of-function phenotypes in vivo.
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.|Hematology / Medical Oncology Division, Department of Medicine, Weill-Cornell Medical College, New York, NY.
authors.forename: Lukas E
authors.initials: LE
authors.lastname: Dow
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Fisher
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.|Weill-Cornell / Rockefeller / Sloan Kettering Tri-Institutional MD-PhD program, New York, NY.
authors.forename: Kevin P
authors.initials: KP
authors.lastname: O'Rourke
authors.affiliation: Hematology / Medical Oncology Division, Department of Medicine, Weill-Cornell Medical College, New York, NY.
authors.forename: Ashlesha
authors.initials: A
authors.lastname: Muley
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.|Gerstner Sloan Kettering Graduate School of Biomedical Sciences, New York, NY.
authors.forename: Edward R
authors.initials: ER
authors.lastname: Kastenhuber
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Geulah
authors.initials: G
authors.lastname: Livshits
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Darjus F
authors.initials: DF
authors.lastname: Tschaharganeh
authors.affiliation: Bioinformatics Core facility, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Nicholas D
authors.initials: ND
authors.lastname: Socci
authors.affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY.|Howard Hughes Medical Institute, Memorial Sloan Kettering Cancer Center, New York, NY.
authors.forename: Scott W
authors.initials: SW
authors.lastname: Lowe
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D016678:Genome / Q000235:genetics*; D051379:Mice; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D011995:Recombination, Genetic / Q000235:genetics