A yeast optogenetic toolkit (yOTK) for gene expression control in Saccharomyces cerevisiae

Citations Over TimeTop 10% of 2019 papers

Abstract

Optogenetic tools for controlling gene expression are ideal for tuning synthetic biological networks due to the exquisite spatiotemporal control available with light. Here we develop an optogenetic system for gene expression control integrated with an existing yeast toolkit allowing for rapid, modular assembly of light-controlled circuits in the important chassis organism Saccharomyces cerevisiae. We reconstitute activity of a split synthetic zinc-finger transcription factor (TF) using light-induced dimerization mediated by the proteins CRY2 and CIB1. We optimize function of this split TF and demonstrate the utility of the toolkit workflow by assembling cassettes expressing the TF activation domain and DNA-binding domain at different levels. Utilizing this TF and a synthetic promoter we demonstrate that light intensity and duty cycle can be used to modulate gene expression over the range currently available from natural yeast promoters. This study allows for rapid generation and prototyping of optogenetic circuits to control gene expression in S. cerevisiae.

Related Papers

• → Non-viral and viral delivery systems for CRISPR-Cas9 technology in the biomedical field(2017)52 cited
• → Cas9 Cuts and Consequences; Detecting, Predicting, and Mitigating CRISPR/Cas9 On‐ and Off‐Target Damage(2020)20 cited
• → [Research progress of CRISPR-Cas9 system for gene therapy].(2016)6 cited
• → Prediction of off-target effects of the CRISPR/Cas9 system for design of sgRNA(2020)
• → The CRISPR/Cas9 system forms a condensate in the yeast nucleus(2023)