Table14_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.

猪克隆受精卵的冷冻保存对生物技术与生物医学研究具有重要意义，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，获取玻璃化冷冻猪克隆受精卵发育至2-细胞期、4-细胞期及囊胚期的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-细胞期的167个差异表达（differentially expressed，DE）mRNA与516个差异表达lncRNA，4-细胞期的469个差异表达mRNA与565个差异表达lncRNA，以及囊胚期的389个差异表达mRNA与816个差异表达lncRNA。功能富集分析结果显示，胚胎发育过程中的差异表达mRNA参与了细胞周期、细胞增殖、凋亡、代谢等多种调控机制。此外，三个胚胎阶段差异表达lncRNA的靶基因同样富集于诸多关键基因本体（Gene Ontology，GO）条目及通路，例如“防御响应”“亚油酸代谢过程”“胚胎轴特化”“蛋白质类泛素化负调控”等。综上，本研究提供了玻璃化冷冻猪克隆受精卵不同发育阶段的mRNA与lncRNA综合转录组数据，有助于进一步解析受损胚胎发育的潜在冷冻损伤机制。