ICAR- National Dairy Research Institute, Karnal, India.
Vet Res Commun. 2024 Aug;48(4):2457-2475. doi: 10.1007/s11259-024-10419-8. Epub 2024 Jun 3.
Somatic cell nuclear transfer (SCNT) is a very important reproductive technology with many diverse applications, such as fast multiplication of elite animals, the production of transgenic animals and embryonic stem (ES) cells. However, low cloning efficiency, a low live birth rate and the abnormally high incidence of abnormalities in the offspring born are attributed to incomplete or aberrant nuclear reprogramming. In SCNT embryos, the aberrant expression pattern of the genes throughout embryonic development is responsible for the incomplete nuclear reprogramming. The present study was carried out to identify the differential gene expression (DEGs) profile and molecular pathways of the SCNT and IVF embryos at different developmental stages (2 cell, 8 cell and blastocyst stages). In the present study, 1164 (2 cell), 1004 (8 cell) and 530 (blastocyst stage) DEGs were identified in the SCNT embryos as compared to IVF embryos. In addition, several genes such as ZEB1, GDF1, HSF5, PDE3B, VIM, TNNC, HSD3B1, TAGLN, ITGA4 and AGMAT were affecting the development of SCNT embryos as compared to IVF embryos. Further, Gene Ontology (GO) and molecular pathways analysis suggested, SCNT embryos exhibit variations compared to their IVF counterparts and affected the development of embryos throughout the different developmental stages. Apart from this, q-PCR analysis of the GDF1, TMEM114, and IGSF22 genes were utilized to validate the RNA-seq data. These findings contribute valuable insights about the different genes and molecular pathways underlying SCNT embryo development and offer crucial information for improving SCNT efficiency.
体细胞核移植(Somatic Cell Nuclear Transfer,SCNT)是一项非常重要的生殖技术,具有许多不同的应用,如快速繁殖优秀动物、生产转基因动物和胚胎干细胞(Embryonic Stem Cells,ES 细胞)。然而,低克隆效率、低活产率和出生后代异常率高归因于不完全或异常的核重编程。在 SCNT 胚胎中,胚胎发育过程中基因的异常表达模式导致核重编程不完全。本研究旨在鉴定不同发育阶段(2 细胞、8 细胞和囊胚阶段)的 SCNT 和 IVF 胚胎的差异基因表达(Differential Gene Expression,DEG)谱和分子途径。在本研究中,与 IVF 胚胎相比,SCNT 胚胎在 2 细胞、8 细胞和囊胚阶段分别鉴定出 1164 个(2 细胞)、1004 个(8 细胞)和 530 个(囊胚阶段)差异表达基因。此外,与 IVF 胚胎相比,ZEB1、GDF1、HSF5、PDE3B、VIM、TNNC、HSD3B1、TAGLN、ITGA4 和 AGMAT 等几个基因也影响了 SCNT 胚胎的发育。此外,基因本体论(Gene Ontology,GO)和分子途径分析表明,与 IVF 胚胎相比,SCNT 胚胎表现出不同的变化,并影响不同发育阶段胚胎的发育。除此之外,还利用 GDF1、TMEM114 和 IGSF22 基因的 q-PCR 分析来验证 RNA-seq 数据。这些发现为 SCNT 胚胎发育的不同基因和分子途径提供了有价值的见解,并为提高 SCNT 效率提供了重要信息。