RGD Reference Report - Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform. - Rat Genome Database

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Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform.

Authors: Yoshimi, K  Kaneko, T  Voigt, B  Mashimo, T 
Citation: Yoshimi K, etal., Nat Commun. 2014 Jun 26;5:4240. doi: 10.1038/ncomms5240.
RGD ID: 11040972
Pubmed: PMID:24967838   (View Abstract at PubMed)
PMCID: PMC4083438   (View Article at PubMed Central)
DOI: DOI:10.1038/ncomms5240   (Journal Full-text)

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 x 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.



Objects referenced in this article
Strain F344-AsipATyrCKitH/Kyo null Rattus norvegicus
Strain F344-TyrCKitH/Kyo null Rattus norvegicus

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