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作为人类胚胎干细胞中主要编辑的大小依赖细胞决定因素的MutSα和MutSβ

Ju-Chan Park¹, Yun-Jeong Kim¹, Jun Hee Han²

¹College of Pharmacy, Seoul National University, Seoul, Republic of Korea.

Molecular therapy. Nucleic acids

|June 22, 2023

概括

DNA不匹配修复蛋白 MutSα 和 MutSβ 在人类多能干细胞 (hPSC) 中显著影响主要编辑器 (PE) 效率. 破坏这些蛋白质可以提高PE的结果,为hPSC中的基因组编辑提供了新的策略.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
干细胞生物学干细胞生物学

背景情况:

人类多能干细胞 (hPSCs) 对疾病建模和再生医学至关重要.
对于hPSCs,需要高效的基因组编辑工具,但它们的独特特性会影响工具的效率.
主编辑器 (PEs) 提供精确的基因组编辑,但需要在hPSCs中进行优化.

研究的目的:

研究DNA不匹配修复 (MMR) 蛋白质在hPSCs中确定主要编辑器 (PE) 效率中的作用.
描述MMR成分MutSα和MutSβ对PE结果的大小依赖的影响.

主要方法:

为关键的MMR蛋白MSH2,MSH3和MSH6.6生成的同卵性淘汰 (KO) hPSCs.
在野生类型和缺乏MMR的hPSC中,评估了各种编辑大小的PE效率.

主要成果:

MutSα和MutSβ复合体以一种取决于所需编辑的大小的方式决定PE效率.
影响MutSα和MutSβ的MSH2 KO,将PE效率提高了50倍,从基数不匹配到10个基数对的编辑.
MSH6 KO (缺少MutSα) 提高了对1-3个基对编辑的效率,而MSH3 KO (缺少MutSβ) 提高了对3-10个基对编辑的效率.

结论:

DNA不匹配修复 (MMR) 途径是hPSCs中主要编辑效率的关键决定因素.
在调节PE结果方面,MutSα和MutSβ表现出不同的,大小依赖的作用.
针对MMR组件提供了一种策略,以提高hPSCs的基因组编辑精度和效率.

关键词:

修复DNA损伤的修复这是一个MMRMMR.MT:RNA/DNA编辑;RNA/DNA编辑;这就是MUTSα.这就是MutsSββ.基因组编辑基因组编辑人类胚胎干细胞人类多能干细胞干细胞总编辑总编辑总编辑

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