High throughput sequencing identifies an imprinted gene, Grb10, associated with the pluripotency state in nuclear transfer embryonic stem cells.
High throughput sequencing identifies an imprinted gene, Grb10, associated with the pluripotency state in nuclear transfer embryonic stem cells.
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高通量测序鉴定出与核移植胚胎干细胞的多能性状态相关的印记基因 Grb10
DOI：
10.18632/oncotarget.17185
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发表时间：
2017-07-18
期刊：
影响因子：
--
通讯作者：
Han Z
中科院分区：
文献类型：
--
作者：
Li H;Gao S;Huang H;Liu W;Huang H;Liu X;Gao Y;Le R;Kou X;Zhao Y;Kou Z;Li J;Wang H;Zhang Y;Wang H;Cai T;Sun Q;Gao S;Han Z
体细胞核移植和转录因子介导的重编程是体细胞重编程的两种广泛应用的技术。完全重编程的核移植胚胎干细胞和诱导的多能干细胞都具有再生医学的潜力，而干细胞多能性状态的评估对这些应用至关重要。以往的报道表明，Dlk1-dio3区域与诱导的多能干细胞的多能性有关，与核移植、胚胎干细胞和胚胎干细胞相比，不完全的体细胞重编程导致诱导的多潜能干细胞的基因组5-甲基胞嘧啶水平异常升高。在这项研究中，我们比较了完全同基因的核移植胚胎干细胞中Dlk1-dio3区域的多能性相关基因Rian和Gtl2，并诱导了具有相同基因组插入的多能性干细胞。我们还评估了5-甲基胞嘧啶和5-羟甲基胞嘧啶的水平，并在这些细胞中进行了高通量测序。我们的结果表明，与诱导多能干细胞多能性相关的Dlk1-dio3区域的Rian和Gtl2与核移植胚胎干细胞中的基因没有相关性，完全和部分重编程核移植胚胎干细胞与诱导多能干细胞之间的5-甲基胞嘧啶和5-羟甲基胞嘧啶水平没有显著差异。通过同源比较，我们的研究首次发现Grb10与核移植胚胎干细胞的多能性状态有关。
Somatic cell nuclear transfer and transcription factor mediated reprogramming are two widely used techniques for somatic cell reprogramming. Both fully reprogrammed nuclear transfer embryonic stem cells and induced pluripotent stem cells hold potential for regenerative medicine, and evaluation of the stem cell pluripotency state is crucial for these applications. Previous reports have shown that the Dlk1-Dio3 region is associated with pluripotency in induced pluripotent stem cells and the incomplete somatic cell reprogramming causes abnormally elevated levels of genomic 5-methylcytosine in induced pluripotent stem cells compared to nuclear transfer embryonic stem cells and embryonic stem cells. In this study, we compared pluripotency associated genes Rian and Gtl2 in the Dlk1-Dio3 region in exactly syngeneic nuclear transfer embryonic stem cells and induced pluripotent stem cells with same genomic insertion. We also assessed 5-methylcytosine and 5-hydroxymethylcytosine levels and performed high-throughput sequencing in these cells. Our results showed that Rian and Gtl2 in the Dlk1-Dio3 region related to pluripotency in induced pluripotent stem cells did not correlate with the genes in nuclear transfer embryonic stem cells, and no significant difference in 5-methylcytosine and 5-hydroxymethylcytosine levels were observed between fully and partially reprogrammed nuclear transfer embryonic stem cells and induced pluripotent stem cells. Through syngeneic comparison, our study identifies for the first time that Grb10 is associated with the pluripotency state in nuclear transfer embryonic stem cells.
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