Imperial College London
Alternate

Professor Mark Isalan - Deputy Head of Department

Faculty of Natural Sciences, Department of Life Sciences

Professor of Synthetic Biology
 
 
 
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Contact

 

+44 (0)20 7594 6482m.isalan

 
 
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Location

 

509Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kogenaru:2018:10.1021/acssynbio.7b00302,
author = {Kogenaru, M and Isalan, M},
doi = {10.1021/acssynbio.7b00302},
journal = {ACS Synthetic Biology},
pages = {1496--1506},
title = {Drug-inducible control of lethality genes: a low background destabilizing domain architecture applied to the Gal4-UAS system in Drosophila},
url = {http://dx.doi.org/10.1021/acssynbio.7b00302},
volume = {7},
year = {2018}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Destabilizing domains (DDs) are genetic tags that conditionally control the level of abundance of proteins-of-interest (POI) with specific stabilizing small-molecule drugs, rapidly and reversibly, in a wide variety of organisms. The amount of the DD-tagged fusion protein directly impacts its molecular function. Hence, it is important that the background levels be tightly regulated in the absence of any drug. This is especially true for classes of proteins that function at extremely low levels, such as lethality genes involved in tissue development and certain transcriptional activator proteins. Here, we establish the uninduced background and induction levels for two widely used DDs (FKBP and DHFR) by developing an accurate quantification method. We show that both DDs exhibit functional background levels in the absence of a drug, but each to a different degree. To overcome this limitation, we systematically test a double architecture for these DDs (DD-POI-DD) that completely suppresses the protein’s function in an uninduced state, while allowing tunable functional levels upon adding a drug. As an example, we generate a drug-stabilizable Gal4 transcriptional activator with extremely low background levels. We show that this functions in vivo in the widely used Gal4-UAS bipartite expression system in Drosophila melanogaster. By regulating a cell death gene, we demonstrate that only the low background double architecture enables tight regulation of the lethal phenotype in vivo. These improved tools will enable applications requiring exceptionally tight control of protein function in living cells and organisms.
AU  - Kogenaru,M
AU  - Isalan,M
DO  - 10.1021/acssynbio.7b00302
EP  - 1506
PY  - 2018///
SN  - 2161-5063
SP  - 1496
TI  - Drug-inducible control of lethality genes: a low background destabilizing domain architecture applied to the Gal4-UAS system in Drosophila
T2  - ACS Synthetic Biology
UR  - http://dx.doi.org/10.1021/acssynbio.7b00302
UR  - https://pubs.acs.org/doi/10.1021/acssynbio.7b00302
UR  - http://hdl.handle.net/10044/1/59919
VL  - 7
ER  -
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