Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes.
Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes.
复制标题
小鼠zygotes中CRISPR/Cas9 RNA引起的体细胞镶嵌和等位基因复杂性。
DOI：
10.1016/j.ydbio.2014.06.017
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发表时间：
2014-09-01
影响因子：
2.7
通讯作者：
Behringer, Richard R.
中科院分区：
文献类型：
--
作者：
Yen, Shuo-Ting;Zhang, Min;Deng, Jian Min;Usman, Shireen J.;Smith, Chad N.;Parker-Thornburg, Jan;Swinton, Paul G.;Martin, James F.;Behringer, Richard R.
酪氨酸酶是黑色素生成的限速酶。在小鼠和其他动物中，酪氨酸酶基因（Tyr）中的纯合无效突变导致色素沉着的缺乏，即白化病。在这里，我们使用CRISPR/Cas9系统通过受精卵注射两种单向导RNA和Cas9 RNA在Tyr基因座中产生单等位基因和双等位基因无效突变。注射到C57 BL/6 N野生型胚胎中导致一个完全白化病创始人携带两种不同的Tyr突变。此外，三个色素沉着马赛克和完全着色的同窝出生的传递新的突变体Tyr等位基因的测试杂交与白化病的后代。注射到Tyr杂合子（B6 CBAF 1/J × FVB/NJ）合子中导致产生大量白化病和具有分级范围的白化病嵌合体的小鼠。深度测序显示，大多数白化病人和嵌合体都有两个以上的新突变等位基因。这些视觉表型和分子基因型突出了在小鼠中通过受精卵注射的CRISPR/Cas9介导的诱变期间针对靶基因发生的创始者中的体细胞嵌合现象和等位基因复杂性。
Tyrosinase is the rate-limiting enzyme for the production of melanin pigmentation. In the mouse and other animals, homozygous null mutations in the Tyrosinase gene (Tyr) result in the absence of pigmentation, i.e. albinism. Here we used the CRISPR/Cas9 system to generate mono- and bi-allelic null mutations in the Tyr locus by zygote injection of two single-guide and Cas9 RNAs. Injection into C57BL/6N wild-type embryos resulted in one completely albino founder carrying two different Tyr mutations. In addition, three pigmentation mosaics and fully pigmented littermates were obtained that transmitted new mutant Tyr alleles to progeny in test crosses with albinos. Injection into Tyr heterozygous (B6CBAF1/J × FVB/NJ) zygotes resulted in the generation of numerous albinos and also mice with a graded range of albino mosaicism. Deep sequencing revealed that the majority of the albinos and the mosaics had more than two new mutant alleles. These visual phenotypes and molecular genotypes highlight the somatic mosaicism and allele complexity in founders that occurs for targeted genes during CRISPR/Cas9-mediated mutagenesis by zygote injection in mice.
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影响因子：
1.5
作者：
Nakayama, Takuya;Fish, Margaret B.;Fisher, Marilyn;Oomen-Hajagos, Jamina;Thomsen, Gerald H.;Grainger, Robert M.
通讯作者：
Grainger, Robert M.
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
4.6
作者：
Horii T;Arai Y;Yamazaki M;Morita S;Kimura M;Itoh M;Abe Y;Hatada I
通讯作者：
Hatada I
影响因子：
1.5
作者：
Blitz, Ira L.;Biesinger, Jacob;Xie, Xiaohui;Cho, Ken W. Y.
通讯作者：
Cho, Ken W. Y.
影响因子：
5.1
作者：
Belhaj K;Chaparro-Garcia A;Kamoun S;Nekrasov V
通讯作者：
Nekrasov V