Disruption of FGF5 in Cashmere Goats Using CRISPR/Cas9 Results in More Secondary Hair Follicles and Longer Fibers.
Disruption of FGF5 in Cashmere Goats Using CRISPR/Cas9 Results in More Secondary Hair Follicles and Longer Fibers.
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使用 CRISPR/Cas9 破坏绒山羊中的 FGF5 会产生更多的次生毛囊和更长的纤维。
DOI：
10.1371/journal.pone.0164640
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Chen Y
中科院分区：
文献类型：
--
作者：
Wang X;Cai B;Zhou J;Zhu H;Niu Y;Ma B;Yu H;Lei A;Yan H;Shen Q;Shi L;Zhao X;Hua J;Huang X;Qu L;Chen Y
精确基因工程加速了农业和生物医学牲畜的遗传改良。我们最近报道了我们使用CRISPR/Cas9系统通过显微注射Cas9 mRNA和靶向山羊胚胎中的MMP 3和FGF 5基因的sgRNA成功地生产基因修饰的山羊。通过研究基因修饰对Cas9介导的山羊表型的影响，我们在本文中证明了这种涉及FGF 5破坏的方法的效用导致Cas9介导的山羊中第二毛囊数量增加和纤维长度增加，这表明将产生更多的羊绒。使用H&E和免疫组织化学染色、定量PCR和蛋白质印迹技术表征基因组修饰的影响。这些结果表明，FGF 5的破坏诱导的基因修饰发生在形态和遗传水平。我们进一步表明，敲除等位基因可能能够生殖系传播，这是山羊种群扩张所必需的。这些结果为使用CRISPR/Cas9方法产生显示相应突变表型的基因修饰山羊的优点提供了充分的证据。
Precision genetic engineering accelerates the genetic improvement of livestock for agriculture and biomedicine. We have recently reported our success in producing gene-modified goats using the CRISPR/Cas9 system through microinjection of Cas9 mRNA and sgRNAs targeting the MSTN and FGF5 genes in goat embryos. By investigating the influence of gene modification on the phenotypes of Cas9-mediated goats, we herein demonstrate that the utility of this approach involving the disruption of FGF5 results in increased number of second hair follicles and enhanced fiber length in Cas9-mediated goats, suggesting more cashmere will be produced. The effects of genome modifications were characterized using H&E and immunohistochemistry staining, quantitative PCR, and western blotting techniques. These results indicated that the gene modifications induced by the disruption of FGF5 had occurred at the morphological and genetic levels. We further show that the knockout alleles were likely capable of germline transmission, which is essential for goat population expansion. These results provide sufficient evidences of the merit of using the CRISPR/Cas9 approach for the generation of gene-modified goats displaying the corresponding mutant phenotypes.
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DOI：
10.1073/pnas.1310478110
发表时间：
2013-10-08
影响因子：
11.1
作者：
Tan, Wenfang;Carlson, Daniel F.;Fahrenkrug, Scott C.
通讯作者：
Fahrenkrug, Scott C.
影响因子：
10.5
作者：
Kuramochi-Miyagawa, Satomi;Watanabe, Toshiaki;Nakano, Toru
通讯作者：
Nakano, Toru
影响因子：
64.5
作者：
HEBERT, JM;ROSENQUIST, T;MARTIN, GR
通讯作者：
MARTIN, GR
影响因子：
11.8
作者：
Ablain J;Durand EM;Yang S;Zhou Y;Zon LI
通讯作者：
Zon LI
成纤维细胞生长因子5-short (FGF5s)抑制绒山羊初级和次级毛囊真皮乳头细胞中FGF5的活性
影响因子：
3.5
作者：
He, Xiaolin;Chao, Yuan;Chen, Yulin
通讯作者：
Chen, Yulin