Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform.
Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR-Cas platform.
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DOI：
10.1038/ncomms5240
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发表时间：
2014-06-26
影响因子：
16.6
通讯作者：
Mashimo, T.
中科院分区：
文献类型：
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作者：
Yoshimi, K.;Kaneko, T.;Voigt, B.;Mashimo, T.
细菌CRISPR/Cas系统已被证明是一种在各种生物体中有效的基因靶向工具。在这里，我们使用CRISPR/CA对大鼠进行准确和高效的基因组编辑。合成的嵌合引导RNAs(GRNAs)区分大鼠胚胎成纤维细胞的单核苷酸多态(SNP)差异，允许对(F344×DA)F1杂交胚胎的显性表型进行等位基因特异性基因组编辑。有趣的是，最初由等位基因特异性gRNA评估的目标等位基因，是通过同源染色体之间的等位基因转换来修复的。使用单链寡核苷酸，我们恢复了三种隐性表型：通过SNP交换的白化表型；通过整合19-bp DNA片段的非刺鼠表型；通过消除7,098-bp插入DNA片段的帽状表型，进化地源自内源性逆转录病毒。CRISPR/Cas系统在体内的成功应用证实了它作为基因工程工具在建立人类疾病动物模型方面的重要性及其在基因治疗中的潜在应用。细菌CRISPR-Cas系统越来越多地用于动物模型的基因组编辑。在这里，作者利用这个系统来定位和编辑大鼠特定的毛色等位基因，并展示了这项技术在创建基因工程动物模型方面的潜力。
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. The bacterial CRISPR–Cas system is increasingly used for genome editing in animal models. Here the authors utilize this system to target and edit specific coat colour alleles in rats and demonstrate the potential of this technology for the creation of genetically engineered animal models.
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影响因子：
48
作者：
Mali, Prashant;Esvelt, Kevin M.;Church, George M.
通讯作者：
Church, George M.
影响因子：
48
作者：
Brown, Andrew J.;Fisher, Daniel A.;Cui, Xiaoxia
通讯作者：
Cui, Xiaoxia
影响因子：
4.6
作者：
Mashimo, Tomoji;Kaneko, Takehito;Sakuma, Tetsushi;Kobayashi, Junya;Kunihiro, Yayoi;Voigt, Birger;Yamamoto, Takashi;Serikawa, Tadao
通讯作者：
Serikawa, Tadao
影响因子：
46.9
作者：
Cho, Seung Woo;Kim, Sojung;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo
影响因子：
16.8
作者：
通讯作者：
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