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Updated: Jun 1, 2025

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

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Published on: February 26, 2019

在性Cas9中分子适应的in-silico研究

Anisha Debnath¹, Aveepsa Sengupta¹, Sujata Rudrapal¹

¹Microbial Adaptation Laboratory, Department of Microbiology, Tripura University (A Central University), Agartala 799022, India.

Letters in applied microbiology

|January 20, 2025

概括

来自喜欢盐的细菌的CRISPR-Cas9蛋白质,如Salicibibacter cibi,表现出结构性适应,以求生存. 这些包括更多的酸性残留物和无序区域,增强盐水环境中的稳定性,以应对潜在的极端条件基因组编辑.

科学领域:

分子生物学分子生物学
结构生物学结构生物学
基因组学就是基因组学.

背景情况:

CRISPR-Cas9系统是一个强大的基因组编辑工具.
了解其在极端环境中的适应性对于扩大其应用至关重要.
型细菌在高盐条件下壮成长,对蛋白质结构和功能提出了独特的挑战.

研究的目的:

为了研究CRISPR-Cas9系统在型细菌中的结构性适应.
分析来自Salicibibacter cibi的Cas9蛋白质,以寻找与其盐水环境相关的独特特征.
探索性Cas9在极端条件下生物技术应用的潜力.

主要方法:

对Cas9蛋白序列的生物信息分析 (ExPASy ProtParam,自然失调区域的预测器,InterPro,WebLogo).
使用AlphaFold进行蛋白质结构建模,使用PROCHECK进行质量评估.
蛋白质表面,氨基酸和静电潜力的可视化 (PyMOL,APBS服务器,UCSF仿真体).

主要成果:

性Cas9蛋白具有较高的酸性残留物,有助于盐水条件下的稳定性和水分.
在性Cas9蛋白中观察到本质上有障碍的区域增加.
静电电位图显示,S. cibi Cas9的表面电荷极为负,这对于适应富含盐的环境至关重要.

结论:

结构和功能上的适应使得CRISPR-Cas9能够在高盐环境中有效地发挥作用.
性Cas9的独特特性,如增强的稳定性和负面电荷,是其生存的关键.
这些发现表明,性Cas9在极端环境的基因组编辑技术中有潜力.

关键词:

沙利西比巴克特 (Salicibibacter cibi) 是一种植物.在CRISPRCas系统中.类的Cas9是类的蛋白质的稳定性蛋白质的稳定性对盐的耐受性有所不同.盐耐受性机制的作用.

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