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J Comput Biol. 2011 Feb;18(2):195-203. doi: 10.1089/cmb.2010.0208.

Design and construction of a versatile synthetic network for bistable gene expression in mammalian systems.

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1
Department of Engineering Mathematics, University of Bristol, Bristol, United Kingdom.

Abstract

We constructed and modeled a novel synthetic network which may be able to exhibit bistable expression of a reporter gene in mammalian cells. This network is based on an aptamer-fused short-hairpin RNA (shRNA) directed against a single mRNA encoding both a EGFP reporter gene and the repressor tTR-KRAB, which, in turn, represses transcription of the shRNA. The activity of the shRNA can be controlled by an inducer molecule (theophylline) which prevents the aptamer-fused shRNA to be properly processed. Repression of the tTR-KRAB can be relieved by treatment with doxycyline. This reciprocal negative feed-back loop can exhibit a bistable response, as shown through the mathematical analysis performed here. Specifically, the network can be controlled to induce sustained expression of a shRNA, or the reporter gene, with a transient input of two different inducer molecules.

PMID:
21314458
DOI:
10.1089/cmb.2010.0208
[Indexed for MEDLINE]
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