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Construction of Gene Regulatory Networks using Synthetic Guide RNAs.

Introduction

Despite the fundamental role transcription factors play, they place many constraints on experimental designs due to limited types available. Synthetic guide RNA can be modulated to match existing promoters, allowing for targeted regulation without the need for DNA manipulation. Inspired by Gander’s study, “Digital logic circuits in yeast with CRISPR-dCas9 NOR gates”, we have decided to build a NOR gate, whose output is controlled via inducible production of gRNA-complexed dCas9. First, we will create four different strains of yeast, each representing a stationary state. The system has two different gRNA; the expression of either results in the inhibition or transcription of yeGFP. Finally, we will create another strain containing a NOR gate, whose different output states can be modified by controlling the production of both gRNAs using different inducible promoters. Thus, our project verifies regulated recruitment using synthetic guide RNA as a viable alternative to conventional transcriptional regulatory modules.