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Transcriptomic Features of Bovine Blastocysts Derived by Somatic Cell Nuclear Transfer

机译:体细胞核转移衍生的牛囊胚的转录组学特征

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摘要

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.
机译:重编程不完全发生在大多数体细胞核移植(SCNT)胚胎中,这会导致发育上重要的基因失调,进而导致胚胎功能障碍和致死性。在这里,我们检查了通过体外受精(IVF)和SCNT衍生的单个牛胚囊的转录组谱。不同类型的供体细胞,卵丘细胞和耳皮成纤维细胞,分别用于衍生cSCNT和fSCNT胚泡。 SCNT胚泡平均表达13,606个基因,类似于IVF(13,542)。相关分析发现,cSCNT和fSCNT胚泡组均具有与IVF组不同的转录组特征,其中cSCNT转录组比fSCNT更接近IVF。基因表达分析确定了两个SCNT组中56个表达不足的基因和78个表达过度的基因。在fSCNT胚泡中发现一个在18号染色体上带有锌指蛋白家族基因的400 kb的基因座被协同下调,表现出抗重编程区的特征。探查对胚泡发育重要的不同类别的基因后发现,与滋养外胚层发育有关的基因常常被低估,而编码表观遗传修饰子的基因在SCNT胚囊中往往被过度代表。我们鉴定抗重编程的差异表达基因的努力可以帮助将重编程易错基因座定位到基因组上,并阐明如何处理重编程的随机事件以提高克隆效率。

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