复杂疾病小鼠模型构建新策略：类精子单倍体胚胎干细胞介导的半克隆技术

doi:10.12300/j.issn.1674-5817.2021.143

摘要/Abstract

摘要： 类精子单倍体胚胎干细胞是源自小鼠孤雄囊胚中的一种新型单倍体胚胎干细胞,仅含有父源遗传物质,性染色体为X染色体,能在体外长期自我更新、增殖或诱导分化,并可以利用CRISPR系统进行单基因或多基因的编辑,能替代精子使卵母细胞受精。区别于原核注射、卵胞质内注射获得嵌合体或利用四倍体补偿技术等传统构建基因修饰小鼠模型的方法,通过将基因编辑后的类精子单倍体胚胎干细胞注射到卵母细胞中,可以高效稳定地一步获得基因型确定的半克隆小鼠,不会存在嵌合现象,原代小鼠即可用于研究。基于类精子单倍体胚胎干细胞获得的多基因精准编辑的复杂疾病小鼠模型,可以在生物个体水平上阐述多个基因协同互作的效应,从而更充分地模拟复杂的、可能受多个基因影响的人类疾病的各种病理特征,以挖掘新的诊断和治疗方法。

关键词: 单倍体, 类精子单倍体胚胎干细胞, CRISPR/Cas9, 小鼠疾病模型

Abstract: The development of haploid genetics has motivated studies on genome evolution and function, especially the technological advancements in recent years have prompted the birth of culture techniques for mammalian haploid embryonic stem cells (haESCs). Sperm-like haESCs are novel haESCs derived from mouse parthenogenetic blastocysts. Sperm-like haESCs only contain paternal genetic material, and their sex chromosome is the X chromosome. They can self-renew, differentiate, and proliferate in vitro for a long time. Furthermore, editing single or multiple genes using the CRISPR system is possible for sperm-like haESCs, which can replace sperms to fertilize oocytes. In contrast to traditional methods for constructing mouse models, such as pronuclear injection, cytoplasmic injection, and tetraploid complementation, by injecting sperm-like haESCs after gene editing into oocytes, semi-cloned mice with a definitive genotype can be obtained efficiently and stably without chimerism, and primary mice can be used for research. The mouse disease model based on multiple precisely edited genes obtained from sperm-like haESCs can explain the effect of multiple genes synergistic interaction at the level of biological individuals to completely simulate various pathological characteristics of complex human diseases that may be affected by multiple genes, and this model facilitates the exploration of novel diagnostic and therapeutic methods.

Key words: Haploid, Sperm-like haploid embryonic stem cells, CRISPR/Cas9, Mouse disease model

中图分类号:

R-332
Q95-33

赖梭梅, 丁一夫, 李劲松. 复杂疾病小鼠模型构建新策略：类精子单倍体胚胎干细胞介导的半克隆技术[J]. 实验动物与比较医学, 2021, 41(5): 369-383.

LAI Suomei, DING Yifu, LI Jinsong. A New Strategy for Constructing Mouse Models of Complex Diseases: Semi-cloning Technology Based on Sperm-like Haploid Embryonic Stem Cells[J]. Laboratory Animal and Comparative Medicine, 2021, 41(5): 369-383.

图/表 4

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