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提高CRISPRRibonucleoprotein中介精确基因编辑小分子在猪纤维细胞的效率

Yunjing Zhao^1,2, Xinyu Li^1,2, Chang Liu¹

¹Key Laboratory of Animal Cellular and Genetic Engineering of Heilongjiang Province, Northeast Agricultural University, Harbin 150030, China.

Animals : an open access journal from MDPI

|March 13, 2024

概括

小分子通过使用CRISPR-RNP技术在猪细胞中增强精确基因编辑. 单个分子显著提高了同质性导向修复效率,超过了改善基因编辑结果的组合.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
基因工程基因工程基因工程

背景情况:

在牲畜中精确的基因编辑对于农业的进步至关重要.
克里斯普尔-核糖核蛋白 (RNP) 复合体为有针对性的基因修改提供了一个强大的工具.
提高同质导向修复 (HDR) 的效率是精确核酸替代的关键.

研究的目的:

研究小分子在改善猪细胞中CRISPR-RNP介导的基因编辑中的有效性.
确定特定的小分子及其最佳度,以提高HDR效率.
评估单个与联合小分子治疗对基因编辑结果的影响.

主要方法:

猪胎儿纤维细胞 (PFF) 使用CRISPR-Cas9 RNP和单链寡核酸 (ssODN) 进行基因编辑.
各种小分子 (L-189,NU7441,SCR7,L755507,RS-1,Brefeldin A) 在确定最佳度下进行了测试.
基因编辑效率通过PCR放大,测序和对照组的比较分析来评估.

主要成果:

在PFF中确定了小分子活性的最佳度.
与对照组相比,单个小分子提高了HDR中介精确基因编辑效率1.71倍至2.28倍.
小分子组合并没有比单分子治疗提高基因编辑效率.

结论:

特定的单个小分子可以显著提高猪细胞中CRISPR-RNP介导的精确基因编辑效率.
使用单个小分子是一种可行的策略,以改善同质导向修复.
这一发现对在猪身上推进基因工程应用具有重要意义.

关键词:

这就是CRISPR/Cas9的作用.这就是HDR HDR.猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪猪精确的基因编辑.小分子分子小分子

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