体细胞核移植
重编程
克隆(编程)
生物
表观遗传学
体细胞
诱导多能干细胞
染色质
计算生物学
遗传学
细胞生物学
胚胎干细胞
细胞
胚胎
胚泡
计算机科学
DNA
基因
胚胎发生
程序设计语言
作者
Juliano Rodrigues Sangalli,Rafael Vilar Sampaio,Tiago Henrique Camara De,Lawrence C. Smith,Flávio Vieira Meirelles
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 225-244
标识
DOI:10.1007/978-1-0716-3064-8_12
摘要
Cloning by somatic cell Nuclear Transfer (SCNT) is a powerful technology capable of reprograming terminally differentiated cells to totipotency for generating whole animals or pluripotent stem cells for use in cell therapy, drug screening, and other biotechnological applications. However, the broad usage of SCNT remains limited due to its high cost and low efficiency in obtaining live and healthy offspring. In this chapter, we first briefly discuss the epigenetic constraints responsible for the low efficiency of SCNT and current attempts to overcome them. We then describe our bovine SCNT protocol for delivering live cloned calves and addressing basic questions about nuclear reprogramming. Other research groups can benefit from our basic protocol and build up on it to improve SCNT in the future. Strategies to correct or mitigate epigenetic errors (e.g., correcting imprinting loci, overexpression of demethylases, chromatin-modifying drugs) can integrate the protocol described here.
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