title: Pervasive Genotypic Mosaicism in Founder Mice Derived from Genome Editing through Pronuclear Injection.
abstract: Genome editing technologies, especially the Cas9/CRISPR system, have revolutionized biomedical research over the past several years. Generation of novel alleles has been simplified to unprecedented levels, allowing for rapid expansion of available genetic tool kits for researchers. However, the issue of genotypic mosaicism has become evident, making stringent analyses of the penetrance of genome-edited alleles essential. Here, we report that founder mice, derived from pronuclear injection of ZFNs or a mix of guidance RNAs and Cas9 mRNAs, display consistent genotypic mosaicism for both deletion and insertion alleles. To identify founders with greater possibility of transmitting the mutant allele through the germline, we developed an effective germline genotyping method. The awareness of the inherent genotypic mosaicism issue with genome editing will allow for a more efficient implementation of the technologies, and the germline genotyping method will save valuable time and resources.
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada, United States of America.
authors.forename: Daniel
authors.initials: D
authors.lastname: Oliver
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada, United States of America.
authors.forename: Shuiqiao
authors.initials: S
authors.lastname: Yuan
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada, United States of America.
authors.forename: Hayden
authors.initials: H
authors.lastname: McSwiggin
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada, United States of America.
authors.forename: Wei
authors.initials: W
authors.lastname: Yan
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism*; D005260:Female; D018703:Founder Effect; D017353:Gene Deletion; D016678:Genome; D005838:Genotype; D005854:Germ Cells / Q000378:metabolism; D006720:Homozygote; D006801:Humans; D007267:Injections; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000378:metabolism; D008969:Molecular Sequence Data; D009030:Mosaicism; D016254:Mutagenesis, Insertional / Q000235:genetics; D017393:RNA Editing / Q000235:genetics*; D015053:Zygote / Q000378:metabolism
chemical_list: D035683:MicroRNAs