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bioRxiv · 10.1101/092338

An Estradiol-Inducible Promoter Enables Fast, Graduated Control of Gene Expression in Fission Yeast

Ohira, M.·Hendrickson, D. G.·McIsaac, R. S.·Rhind, N.
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Abstract

The fission yeast Schizosaccharomyces pombe lacks a diverse toolkit of inducible promoters for experimental manipulation. Available inducible promoters suffer from slow induction kinetics, limited control of expression levels and/or a requirement for defined growth medium. In particular, no S. pombe inducible promoter systems exhibit a linear dose response, which would allow expression to be tuned to specific levels. We have adapted a fast, orthogonal promoter system with a large dynamic range and a linear dose response, based on {beta}-estradiol-regulated function of the human estrogen receptor, for use in S. pombe. We show that this promoter system, termed Z3EV, turns on quickly, can reach a maximal induction of 20 fold, and exhibits a linear dose response over its entire induction range, with few off target effects. We demonstrate the utility of this system by regulating the mitotic inhibitor Wee1 to create a strain in which cell size is regulated by {beta}-estradiol concentration. This promoter system will be of great utility for experimentally regulating gene expression in fission yeast.

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Ohira, M., Hendrickson, D. G., McIsaac, R. S., Rhind, N.. 2016-12-07. An Estradiol-Inducible Promoter Enables Fast, Graduated Control of Gene Expression in Fission Yeast. https://doi.org/10.1101/092338

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