Pervasive Genotypic Mosaicism in Founder Mice Derived from Genome Editing through Pronuclear Injection

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 issu...

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Bibliographic Details
Published in:PloS one 2015-06, Vol.10 (6), p.e0129457-e0129457
Main Authors: Oliver, Daniel, Yuan, Shuiqiao, McSwiggin, Hayden, Yan, Wei
Format: Article
Language:English
Subjects:
Alleles
Analysis
Animals
Base Sequence
Biology
Cell Nucleus - metabolism
Cost control
CRISPR
CRISPR-Cas Systems - genetics
Deoxyribonucleic acid
DNA
Drosophila
Editing
Engineering
Female
Founder Effect
Gene Deletion
Gene mutation
Genes
Genetic research
Genome
Genome editing
Genomes
Genomics
Genotype
Genotyping
Germ Cells - metabolism
Homozygote
House mouse
Humans
Injection
Injections
Insects
Insertion
Laboratories
Medical research
Medicine
Mice, Inbred C57BL
MicroRNAs - metabolism
Molecular Sequence Data
Mosaicism
Mutagenesis
Mutagenesis, Insertional - genetics
Physiology
RNA Editing - genetics
Target recognition
Zinc finger proteins
Zygote - metabolism
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Description
Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0129457