H3K9me3和H3K27me3是兔体细胞核移植的表观遗传障碍。
H3K9me3 and H3K27me3 are epigenetic barriers to somatic cell nuclear transfer in rabbits.
作者信息
Gao Yuan, Wu Fangfang, Tang Dandan, Chen Xiao, Chen Qian, Chen Ying, Fang Kexin, Chen Wanxin, Du Fuliang, Liu Zhihui
机构信息
Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University Nanjing 210046, Jiangsu, PR China.
Changde First Hospital of Traditional Chinese Medicine, Changde Affiliated Hospital, Hunan University of Chinese Medicine Changde 415000, Hunan, PR China.
出版信息
Am J Transl Res. 2025 Apr 15;17(4):3094-3108. doi: 10.62347/FQYG9348. eCollection 2025.
Somatic cell nuclear transfer (NT) is associated with aberrant changes in epigenetic reprogramming that disrupt embryo development. Here, we characterized the epigenetic patterns of histone 3 lysine 9 trimethylation (H3K9me3) and histone 3 lysine 27 trimethylation (H3K27me3), the repressive histone signature in heterochromatin, in NT rabbit embryos. H3K9me3 modification was higher in all stages of NT vs. fertilized preimplantation embryos, consistent with decreased transcription of the H3K9me3 demethylase in four-cell and eight-cell NT embryos vs. fertilized embryos. H3K27me3 was higher in NT embryos from two-cell stage to the blastocyst, and transcription of the H3K27me3 demethylase was lower in four-cell and eight-cell NT embryos than in fertilized embryos. Cleavage under targets and tagmentation (CUT and Tag) showed enrichment of whole-chromosome H3K9me3 and H3K27me3 in four-cell and eight-cell NT vs. fertilized embryos. Further, eight-cell NT embryos had a higher proportion of H3K9me3 and H3K27me3 sites around the promoter regions of the genome compared to fertilized embryos. The genes associated with H3K9me3 sites near promoter regions were primarily related to the development of tissues and organs, whereas the H3K27me3 enriched regions mainly influenced the development and regulation of the nervous system. Our findings demonstrate that H3K9me3 and H3K27me3 inhibit the expression of genes essential for the development and preimplantation of NT embryos and, thus, are epigenetic barriers to NT reprogramming in rabbits.
体细胞核移植(NT)与表观遗传重编程中的异常变化相关,这些变化会干扰胚胎发育。在此,我们对组蛋白3赖氨酸9三甲基化(H3K9me3)和组蛋白3赖氨酸27三甲基化(H3K27me3)的表观遗传模式进行了表征,这两种修饰是异染色质中的抑制性组蛋白标记,存在于NT兔胚胎中。与受精的植入前胚胎相比,NT胚胎各阶段的H3K9me3修饰水平更高,这与四细胞和八细胞NT胚胎中H3K9me3去甲基化酶的转录水平低于受精胚胎一致。从二细胞阶段到囊胚期,NT胚胎中的H3K27me3水平更高,并且四细胞和八细胞NT胚胎中H3K27me3去甲基化酶的转录水平低于受精胚胎。切割与标签化(CUT和Tag)显示,与受精胚胎相比,四细胞和八细胞NT胚胎中全染色体H3K9me3和H3K27me3富集。此外,与受精胚胎相比,八细胞NT胚胎在基因组启动子区域周围具有更高比例的H3K9me3和H3K27me3位点。启动子区域附近与H3K9me3位点相关的基因主要与组织和器官的发育有关,而H3K27me3富集区域主要影响神经系统的发育和调节。我们的研究结果表明,H3K9me3和H3K27me3抑制了NT胚胎发育和植入前所需基因的表达,因此是兔NT重编程的表观遗传障碍。
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