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| Status |
Public on Dec 13, 2016 |
| Title |
An Estradiol-Inducible Promoter Enables Fast, Graduated Control of Gene Expression in Fission Yeast |
| Organism |
Schizosaccharomyces pombe |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
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 _-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 _-estradiol concentration. This promoter system will be of great utility for experimentally regulating gene expression in fission yeast.
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| |
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| Overall design |
RNA-seq for asynchronous S. pombe cells, +/- beta-estradiol, in biological triplicate
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| |
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| Contributor(s) |
Ohira M, Hendrickson DG, McIsaac RS, Rhind N |
| Citation(s) |
28423198 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 GM098815 |
The Regulation of DNA Replication Kinetics |
UNIV OF MASSACHUSETTS MED SCHOOL |
NICHOLAS R RHIND |
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| |
| Submission date |
Dec 12, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Nick Rhind |
| E-mail(s) |
nick.rhind@umassmed.edu
|
| Phone |
(508) 856-8316
|
| Organization name |
UMass Med School
|
| Department |
Biochemistry and Molecular Pharmalology
|
| Street address |
364 Plantation St.
|
| City |
Worcester |
| State/province |
MA |
| ZIP/Postal code |
01605 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL16192 |
Illumina MiSeq (Schizosaccharomyces pombe) |
|
| Samples (6)
|
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| Relations |
| BioProject |
PRJNA357026 |
| SRA |
SRP094927 |
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