Table7_Transcriptome Analysis of mRNAs and Long Non-Coding RNAs During Subsequent Embryo Development of Porcine Cloned Zygotes After Vitrification.XLSX

Cryopreservation of porcine cloned zygotes has important implications for biotechnology and biomedicine research; however, lower embryo developmental potential remains an urgent problem to be resolved. For exploring the sublethal cryodamages during embryo development, this study was designed to acquire the mRNA and long non-coding RNA (lncRNA) profiles of 2-cells, 4-cells and blastocysts derived from vitrified porcine cloned zygotes using transcriptome sequencing. We identified 167 differentially expressed (DE) mRNAs and 516 DE lncRNAs in 2-cell stage, 469 DE mRNAs and 565 lncRNAs in 4-cell stage, and 389 DE mRNAs and 816 DE lncRNAs in blastocyst stage. Functional enrichment analysis revealed that the DE mRNAs during embryo development were involved in many regulatory mechanisms related to cell cycle, cell proliferation, apoptosis, metabolism and others. Moreover, the target genes of DE lncRNAs in the three embryonic stages were also enriched in many key GO terms or pathways such as “defense response”, “linoleic acid metabolic process”, “embryonic axis specification”, “negative regulation of protein neddylation”, etc., In conclusion, the present study provided comprehensive transcriptomic data about mRNAs and lncRNAs for the vitrified porcine cloned zygotes during different developmental stages, which contributed to further understand the potential cryodamage mechanisms responsible for impaired embryo development.

猪克隆受精卵的低温保存（cryopreservation）对生物技术与生物医学研究具有重要的研究价值与应用意义，但胚胎发育潜能低下仍是亟待解决的紧迫问题。为探明胚胎发育过程中的亚致死冷冻损伤，本研究采用转录组测序技术，获取了玻璃化冷冻猪克隆受精卵来源的2-细胞期、4-细胞期及囊胚的信使RNA（messenger RNA, mRNA）与长链非编码RNA（long non-coding RNA, lncRNA）表达谱。本研究共鉴定得到：2-细胞期存在167个差异表达（differentially expressed, DE）mRNA及516个DE lncRNAs，4-细胞期存在469个DE mRNA与565个长链非编码RNA，囊胚期存在389个DE mRNA与816个DE lncRNAs。功能富集分析表明，胚胎发育过程中的差异表达mRNA参与了细胞周期、细胞增殖、细胞凋亡、代谢等多种调控机制。此外，三个胚胎阶段中差异表达长链非编码RNA的靶基因，亦富集于"防御应答""亚油酸代谢过程""胚胎轴特化""蛋白质Neddylation修饰负调控"等多项关键基因本体（Gene Ontology, GO）术语及信号通路中。综上，本研究为玻璃化冷冻猪克隆受精卵不同发育阶段的信使RNA与长链非编码RNA提供了全面的转录组数据，可为进一步解析胚胎发育受损的潜在冷冻损伤机制提供重要支撑。