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在Saccharomyces cerevisiae中的快速抗体增强平台使用改进的,多样化的CRISPR基编辑器

Andrew P Cazier¹, Olivia M Irvin¹, Lizmarie S Chávez¹

¹School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, United States.

ACS synthetic biology

|October 24, 2023

概括

我们开发了yDBE,一种酵母多样化基编辑器,以加速蛋白质工程. 这种工具可以在体内实现快速,有针对性的DNA多样化,增强针对性进化以改善蛋白质特征.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
合成生物学合成生物学

背景情况:

酵母,特别是*Saccharomyces cerevisiae*,是蛋白质变异选和定向进化的关键平台.
现有的基于酵母的定向进化方法需要提高速度和易用性.

研究的目的:

开发一种新的,快速的,高宽度的酵母DNA多样化工具.
提高使用*Saccharomyces cerevisiae*进行定向进化实验的效率.

主要方法:

开发了yDBE,一种酵母多样化基编辑器,利用CRISPR-dCas9导向的细胞胺脱氨酶进行体内DNA多样化.
通过改进的除氨酶变体和脚手架指导结构,优化了基础编辑器性能.
通过图书馆多样化和细胞分类来改善抗体scFv亲和力,证明了yDBE的有效性.

主要成果:

在100bp窗口中,yDBE的突变率为2.13 × 10^-4替代/bp/代.
成功生成多样化的库并分离出改进的抗体结合剂.
yDBE完全在体内运行,并可针对各种DNA点进行编程.

结论:

yDBE提供了一种强大,快速和有针对性的方法,用于酵母中的DNA多样化.
这种工具可以显著促进广泛的定向进化实验.
yDBE通过简化具有改进功能的蛋白质的开发来增强蛋白质工程能力.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?抗体是一种抗体.基础编辑基础编辑指导进化是指导进化的.合成生物学合成生物学酵母显示器可以显示酵母.

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