Min Byungkuk, Cho Sunwha, Park Jung Sun, Lee Yun-Gyeong, Kim Namshin, Kang Yong-Kook
Development and Differentiation Research Center, KRIBB, 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806, South Korea Department of Functional Genomics, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, 305-350, South Korea.
Development and Differentiation Research Center, KRIBB, 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806, South Korea.
G3 (Bethesda). 2015 Sep 3;5(12):2527-38. doi: 10.1534/g3.115.020016.
Reprogramming incompletely occurs in most somatic cell nuclear transfer (SCNT) embryos, which results in misregulation of developmentally important genes and subsequent embryonic malfunction and lethality. Here we examined transcriptome profiles in single bovine blastocysts derived by in vitro fertilization (IVF) and SCNT. Different types of donor cells, cumulus cell and ear-skin fibroblast, were used to derive cSCNT and fSCNT blastocysts, respectively. SCNT blastocysts expressed 13,606 genes on average, similar to IVF (13,542). Correlation analysis found that both cSCNT and fSCNT blastocyst groups had transcriptomic features distinctive from the IVF group, with the cSCNT transcriptomes closer to the IVF ones than the fSCNT. Gene expression analysis identified 56 underrepresented and 78 overrepresented differentially expressed genes in both SCNT groups. A 400-kb locus harboring zinc-finger protein family genes in chromosome 18 were found coordinately down-regulated in fSCNT blastocysts, showing a feature of reprogramming-resistant regions. Probing into different categories of genes important for blastocyst development revealed that genes involved in trophectoderm development frequently were underrepresented, and those encoding epigenetic modifiers tended to be overrepresented in SCNT blastocysts. Our effort to identify reprogramming-resistant, differentially expressed genes can help map reprogramming error-prone loci onto the genome and elucidate how to handle the stochastic events of reprogramming to improve cloning efficiency.
重编程在大多数体细胞核移植(SCNT)胚胎中不完全发生,这导致发育重要基因的调控异常以及随后的胚胎功能障碍和致死率。在这里,我们检测了通过体外受精(IVF)和SCNT获得的单个牛囊胚的转录组图谱。分别使用不同类型的供体细胞,即卵丘细胞和耳皮肤成纤维细胞,来获得cSCNT和fSCNT囊胚。SCNT囊胚平均表达13,606个基因,与IVF囊胚(13,542个)相似。相关性分析发现,cSCNT和fSCNT囊胚组都具有与IVF组不同的转录组特征,cSCNT的转录组比fSCNT的更接近IVF的转录组。基因表达分析确定了两个SCNT组中56个表达不足和78个表达过度的差异表达基因。在18号染色体上发现一个含有锌指蛋白家族基因的400-kb位点在fSCNT囊胚中协同下调,显示出重编程抗性区域的特征。对囊胚发育重要的不同类别基因进行探究发现,参与滋养外胚层发育的基因在SCNT囊胚中经常表达不足,而编码表观遗传修饰因子的基因往往表达过度。我们识别重编程抗性差异表达基因的工作有助于在基因组上绘制重编程易出错位点,并阐明如何处理重编程的随机事件以提高克隆效率。