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A yeast optogenetic toolkit (yOTK) for gene expression control in Saccharomyces cerevisiae
Author(s) -
Anadirekkun Jidapas My,
Stewart Cameron J.,
Geller Stephanie H.,
Patel Michael T.,
Melendez Justin,
Oakes Benjamin L.,
Noyes Marcus B.,
McClean Megan N.
Publication year - 2020
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27234
Subject(s) - optogenetics , synthetic biology , saccharomyces cerevisiae , computational biology , biology , crispr , gene expression , transcription factor , gene , promoter , yeast , microbiology and biotechnology , genetics , neuroscience
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 .