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通过CRISPR/Cas9介导的胚胎细胞补充来产生外源性脏,以小鼠的Osr1基因为向

Ting-Yu Yeh¹, Ing-Ming Chiu^1,2,3,4, Yu-Jia Huang⁵

¹Doctoral Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung, 402, Taiwan.

Scientific reports

|November 4, 2025

概括

通过胚胎囊补充生成小鼠脏显示出有前途. Osr1淘汰赛和胚胎干细胞注射导致了一些可行的仿真脏,表明器官生成的潜力.

科学领域:

发展生物学发展生物学
再生医学是一种再生医学.
基因编辑技术的技术

背景情况:

胚胎囊补充能够产生外源性小鼠器官,包括胰腺,肺和脏.
以前的补充试验没有成功地拯救宿主动物.

研究的目的:

用两步的胚胎细胞补充策略来产生小鼠仿真脏.
研究Osr1在中管发育和形成中的作用.

主要方法:

用CRISPR/Cas9核糖蛋白 (RNP) 来淘汰小鼠细胞质中的Osr1等位基因.
将表达增强绿色光蛋白 (EGFP) 的小鼠胚胎干细胞 (ES) 注射到胚胎囊中.
用血清化学染色和EGFP表达来评估脏发育和化.

主要成果:

针对Osr1的两个sgRNAs的微注射导致了34%的双基淘汰,导致由于中管发育失败而导致胚胎死亡.
增加sgRNA注射提高了淘汰效率,并使E12.5胚胎中EGFP阳性细胞的中管发展成为可能.
虽然大多数嵌合体胚胎退化,但264个胚胎中有4个存活到产后第二天,在脏和心脏中显示EGFP信号.

结论:

补充Osr1-knockout芽细胞是产生外源性小鼠脏的潜在策略.
需要进一步优化,以提高仿真发育的效率和可行性.

关键词:

胚胎囊补充胚胎囊补充奇梅拉 Chimera 是一个奇梅拉.胚胎干细胞是一种胚胎干细胞.尼弗洛基尼斯发生.美国的奥斯陆.

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In-vitro Mutagenesis

In-vitro Mutagenesis

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