Liu Zhen, Cai Yijun, Liao Zhaodi, Xu Yuting, Wang Yan, Wang Zhanyang, Jiang Xiaoyu, Li Yuzhuo, Lu Yong, Nie Yanhong, Zhang Xiaotong, Li Chunyang, Bian Xinyan, Poo Mu-Ming, Chang Hung-Chun, Sun Qiang
Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, CAS Key Laboratory of Primate Neurobiology, State Key Laboratory of Neuroscience, Chinese Academy of Sciences, Shanghai 200031, China.
Shanghai Research Center for Brain Science and Brain-inspired Technology, Shanghai 200031, China.
Natl Sci Rev. 2019 Jan;6(1):101-108. doi: 10.1093/nsr/nwz003. Epub 2019 Jan 24.
Cloning of macaque monkeys by somatic cell nucleus transfer (SCNT) allows the generation of monkeys with uniform genetic backgrounds that are useful for the development of non-human primate models of human diseases. Here, we report the feasibility of this approach by SCNT of fibroblasts from a macaque monkey (), in which a core circadian transcription factor BMAL1 was knocked out by clustered regularly interspaced short palindromic repeat/Cas9 gene editing (see accompanying paper). Out of 325 SCNT embryos transferred into 65 surrogate monkeys, we cloned five macaque monkeys with mutations in both alleles without mosaicism, with nuclear genes identical to that of the fibroblast donor monkey. Further peripheral blood mRNA analysis confirmed the complete absence of the wild-type transcript. This study demonstrates that the SCNT approach could be used to generate cloned monkeys from fibroblasts of a young adult monkeys and paves the way for the development of macaque monkey disease models with uniform genetic backgrounds.
通过体细胞核移植（SCNT）克隆猕猴能够产生具有一致遗传背景的猴子，这对于人类疾病非人类灵长类动物模型的开发非常有用。在此，我们报告了通过对一只猕猴（）的成纤维细胞进行SCNT来实现这种方法的可行性，其中核心昼夜节律转录因子BMAL1通过成簇规律间隔短回文重复序列/Cas9基因编辑被敲除（见随附论文）。在移植到65只代孕猕猴体内的325个SCNT胚胎中，我们克隆出了五只两个等位基因均有突变且无嵌合体的猕猴，其核基因与成纤维细胞供体猕猴相同。进一步的外周血mRNA分析证实完全不存在野生型转录本。这项研究表明，SCNT方法可用于从成年幼猴的成纤维细胞中产生克隆猴，并为开发具有一致遗传背景的猕猴疾病模型铺平了道路。
