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Article: Engineering a portable riboswitch-LacP hybrid device for two-way gene regulation

TitleEngineering a portable riboswitch-LacP hybrid device for two-way gene regulation
Authors
Issue Date2011
PublisherOxford University Press. The Journal's web site is located at http://nar.oxfordjournals.org/
Citation
Nucleic Acids Research, 2011, v. 39 n. 19 How to Cite?
AbstractRiboswitches are RNA-based regulatory devices that mediate ligand-dependent control of gene expression. However, there has been limited success in rationally designing riboswitches. Moreover, most previous riboswitches are confined to a particular gene and only perform one-way regulation. Here, we used a library screening strategy for efficient creation of ON and OFF riboswitches of lacI on the chromosome of Escherichia coli. We then engineered a riboswitch-LacP hybrid device to achieve portable gene control in response to theophylline and IPTG. Moreover, this device regulated target expression in a 'two-way' manner: the default state of target expression was ON; the expression was switched off by adding theophylline and restored to the ON state by adding IPTG without changing growth medium. We showcased the portability and two-way regulation of this device by applying it to the small RNA CsrB and the RpoS protein. Finally, the use of the hybrid device uncovered an inhibitory role of RpoS in acetate assimilation, a function which is otherwise neglected using conventional genetic approaches. Overall, this work establishes a portable riboswitch-LacP device that achieves sequential OFF-and-ON gene regulation. The two-way control of gene expression has various potential scientific and biotechnological applications and helps reveal novel gene functions. © The Author(s) 2011. Published by Oxford University Press.
Persistent Identifierhttp://hdl.handle.net/10722/137216
ISSN
2015 Impact Factor: 9.202
2015 SCImago Journal Rankings: 7.458
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
Hong Kong Innovation and Technology CommissionITS/148/09
Research Fund for the Control of Infectious Diseases Commissioned Study of Food and Health Bureau of Hong Kong Government
Research Grants CouncilHKU1/CRF/10
HKU201007176077
Funding Information:

Funding for open access charge: An Innovation and Technology Fund (ITS/148/09) from Hong Kong Innovation and Technology Commission, in part by a Research Fund for the Control of Infectious Diseases Commissioned Study of Food and Health Bureau of Hong Kong Government; a Research Grants Council CRF grant (HKU1/CRF/10 to J.-D.H.); a HKU small grant funding (201007176077 to Y.J.).

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorJin, Yen_HK
dc.contributor.authorHuang, JDen_HK
dc.date.accessioned2011-08-26T14:19:04Z-
dc.date.available2011-08-26T14:19:04Z-
dc.date.issued2011en_HK
dc.identifier.citationNucleic Acids Research, 2011, v. 39 n. 19en_HK
dc.identifier.issn0305-1048en_HK
dc.identifier.urihttp://hdl.handle.net/10722/137216-
dc.description.abstractRiboswitches are RNA-based regulatory devices that mediate ligand-dependent control of gene expression. However, there has been limited success in rationally designing riboswitches. Moreover, most previous riboswitches are confined to a particular gene and only perform one-way regulation. Here, we used a library screening strategy for efficient creation of ON and OFF riboswitches of lacI on the chromosome of Escherichia coli. We then engineered a riboswitch-LacP hybrid device to achieve portable gene control in response to theophylline and IPTG. Moreover, this device regulated target expression in a 'two-way' manner: the default state of target expression was ON; the expression was switched off by adding theophylline and restored to the ON state by adding IPTG without changing growth medium. We showcased the portability and two-way regulation of this device by applying it to the small RNA CsrB and the RpoS protein. Finally, the use of the hybrid device uncovered an inhibitory role of RpoS in acetate assimilation, a function which is otherwise neglected using conventional genetic approaches. Overall, this work establishes a portable riboswitch-LacP device that achieves sequential OFF-and-ON gene regulation. The two-way control of gene expression has various potential scientific and biotechnological applications and helps reveal novel gene functions. © The Author(s) 2011. Published by Oxford University Press.en_HK
dc.languageengen_US
dc.publisherOxford University Press. The Journal's web site is located at http://nar.oxfordjournals.org/en_HK
dc.relation.ispartofNucleic Acids Researchen_HK
dc.titleEngineering a portable riboswitch-LacP hybrid device for two-way gene regulationen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0305-1048&volume=39&issue=19, article no. e131&spage=&epage=&date=2011&atitle=Engineering+a+portable+riboswitch-LacP+hybrid+device+for+two-way+gene+regulationen_US
dc.identifier.emailHuang, JD:jdhuang@hkucc.hku.hken_HK
dc.identifier.authorityHuang, JD=rp00451en_HK
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1093/nar/gkr609en_HK
dc.identifier.pmid21803790en_US
dc.identifier.pmcidPMC3201887-
dc.identifier.scopuseid_2-s2.0-80455168324en_HK
dc.identifier.hkuros191797en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-80455168324&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume39en_HK
dc.identifier.issue19en_HK
dc.identifier.eissn1362-4962-
dc.identifier.isiWOS:000296341100006-
dc.publisher.placeUnited Kingdomen_HK
dc.relation.projectProgramming the Second Generation Tumor-targeting Bacteria-
dc.relation.projectDevelopment of a Pan-organism Riboregulator Device-
dc.identifier.scopusauthoridJin, Y=29467538100en_HK
dc.identifier.scopusauthoridHuang, JD=8108660600en_HK
dc.identifier.citeulike9645412-

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