RRC ID 31704
Author Yoshimi K, Kaneko T, Voigt B, Mashimo T.
Title Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform.
Journal Nat Commun
Abstract The bacterial CRISPR/Cas system has proven to be an efficient gene-targeting tool in various organisms. Here we employ CRISPR/Cas for accurate and efficient genome editing in rats. The synthetic chimeric guide RNAs (gRNAs) discriminate a single-nucleotide polymorphism (SNP) difference in rat embryonic fibroblasts, allowing allele-specific genome editing of the dominant phenotype in (F344 × DA)F1 hybrid embryos. Interestingly, the targeted allele, initially assessed by the allele-specific gRNA, is repaired by an interallelic gene conversion between homologous chromosomes. Using single-stranded oligodeoxynucleotides, we recover three recessive phenotypes: the albino phenotype by SNP exchange; the non-agouti phenotype by integration of a 19-bp DNA fragment; and the hooded phenotype by eliminating a 7,098-bp insertional DNA fragment, evolutionary-derived from an endogenous retrovirus. Successful in vivo application of the CRISPR/Cas system confirms its importance as a genetic engineering tool for creating animal models of human diseases and its potential use in gene therapy.
Volume 5
Pages 4240
Published 2014-6-26
DOI 10.1038/ncomms5240
PII ncomms5240
PMID 24967838
PMC PMC4083438
MeSH Agouti Signaling Protein / genetics Alleles Animals CRISPR-Cas Systems* Genetic Engineering / methods* Genetic Therapy / methods Monophenol Monooxygenase / genetics Phenotype Polymorphism, Single Nucleotide Proto-Oncogene Proteins c-kit / genetics RNA Editing* RNA, Guide, Kinetoplastida* Rats
IF 12.121
Times Cited 107
DNA material pT7-gRNA:Tyr^c DR274 (albino) (RDB13010) pT7-gRNA:Tyr^c DR274 (wild) (RDB13011) pT7-gRNA:Asip^a DR274 (RDB13012) pT7-gRNA:kit^h -1 DR274 (RDB13013) pT7-gRNA:kit^h -2-1 DR274 (RDB13014) pT7-gRNA:kit^h -2-2 DR274 (RDB13015).
Rats F344/Stm(strainID=266) DA/Slc(strainID=380) PVG/Seac(strainID=113)
Human and Animal Cells Rat-1(RCB1830)