Orthogonal Modular Gene Repression in Escherichia coli Using Engineered CRISPR/Cas9.

ABSTRACT: The progress in development of synthetic gene circuits has been hindered by the limited repertoire of available transcription factors. Recently, it has been greatly expanded using the CRISPR/Cas9 system. However, this system is limited by its imperfect DNA sequence specificity, leading to potential crosstalk with host genome or circuit components. Furthermore, CRISPR/Cas9-mediated gene regulation is context dependent, affecting the modularity of Cas9-based transcription factors. In this paper we address the problems of specificity and modularity by developing a computational approach for selecting Cas9/gRNA transcription factor/promoter pairs that are maximally orthogonal to each other as well as to the host genome and synthetic circuit components. We validate the method by designing and e

SUBMITTER: Didovyk A 

PROVIDER: S-EPMC5462464 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature