效应器
计算生物学
引导RNA
核糖核酸
合成生物学
清脆的
生物
DNA
计算机科学
遗传学
基因
基因组编辑
细胞生物学
作者
Lukas Oesinghaus,Friedrich C. Simmel
标识
DOI:10.1038/s41467-019-09953-w
摘要
Abstract The CRISPR effector protein Cas12a has been used for a wide variety of applications such as in vivo gene editing and regulation or in vitro DNA sensing. Here, we add programmability to Cas12a-based DNA processing by combining it with strand displacement-based reaction circuits. We first establish a viable strategy for augmenting Cas12a guide RNAs (gRNAs) at their 5′ end and then use such 5′ extensions to construct strand displacement gRNAs (SD gRNAs) that can be activated by single-stranded RNA trigger molecules. These SD gRNAs are further engineered to exhibit a digital and orthogonal response to different trigger RNA inputs—including full length mRNAs—and to function as multi-input logic gates. We also demonstrate that SD gRNAs can be designed to work inside bacterial cells. Using such in vivo SD gRNAs and a DNase inactive version of Cas12a (dCas12a), we demonstrate logic gated transcriptional control of gene expression in E. coli .
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