RRC ID |
31704
|
著者 |
Yoshimi K, Kaneko T, Voigt B, Mashimo T.
|
タイトル |
Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform.
|
ジャーナル |
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.
|
巻・号 |
5
|
ページ |
4240
|
公開日 |
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
|
引用数 |
107
|
WOS 分野
|
GENETICS & HEREDITY
|
リソース情報 |
遺伝子材料 |
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). |
ラット |
F344/Stm(strainID=266)
DA/Slc(strainID=380)
PVG/Seac(strainID=113) |
ヒト・動物細胞 |
Rat-1(RCB1830) |