Show simple item record

dc.contributor.authorMorse, Daniel P.
dc.contributor.authorNevins, Colin E.
dc.contributor.authorAggrey-Fynn, Joana Efua
dc.contributor.authorBravo, Rick J.
dc.contributor.authorPfaeffle, Herman O.I.
dc.contributor.authorLaney, Jess E.
dc.date.accessioned2020-01-07T10:45:22Z
dc.date.available2020-01-07T10:45:22Z
dc.date.issued2018-04en_US
dc.identifier.citationMorse, D. P., Nevins, C. E., Aggrey-Fynn, J. E., Bravo, R. J., Pfaeffle, H.O.I., and Laney, J. E. (2018). Sensitive and specific detection of ligands using engineered riboswitches. Journal of Biotechnology, 272-273, 22-32. doi:10.1016/j.jbiotec.2018.03.002en_US
dc.identifier.issn0168-1656
dc.identifier.urihttps://doi.org/10.1016/j.jbiotec.2018.03.002
dc.identifier.urihttps://hdl.handle.net/11147/7566
dc.description.abstractRiboswitches are RNA elements found in non-coding regions of messenger RNAs that regulate gene expression through a ligand-triggered conformational change. Riboswitches typically bind tightly and specifically to their ligands, so they have the potential to serve as highly effective sensors in vitro. In B. subtilis and other gram-positive bacteria, purine nucleotide synthesis is regulated by riboswitches that bind to guanine. We modified the xpt-pbuX guanine riboswitch for use in a fluorescence quenching assay that allowed us to specifically detect and quantify guanine in vitro. Using this assay, we reproducibly detected as little as 5 nM guanine. We then produced sensors for 2′-deoxyguanosine and cyclic diguanylate (c-diGMP) by appending the P1 stem of the guanine riboswitch to the ligand-binding domains of a 2′-deoxyguanosine riboswitch and a c-diGMP riboswitch. These hybrid sensors could detect 15 nM 2′-deoxyguanosine and 3 nM c-diGMP, respectively. Each sensor retained the ligand specificity of its corresponding natural riboswitch. In order to extend the utility of our approach, we developed a strategy for the in vitro selection of sensors with novel ligand specificity. Here we report a proof-of-principle experiment that demonstrated the feasibility of our selection strategy.en_US
dc.description.sponsorshipUnited States Department of Defense-Defense Threat Reduction Agency (MIPR HDTRA1620511); Office of Naval Research; Chemistry Department of the U.S. Naval Academyen_US
dc.language.isoengen_US
dc.publisherElsevieren_US
dc.relation.isversionof10.1016/j.jbiotec.2018.03.002en_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectBiosensorsen_US
dc.subjectSpecificityen_US
dc.subjectSensitivityen_US
dc.subjectGene expressionen_US
dc.subjectLigandsen_US
dc.subjectRiboswitchesen_US
dc.titleSensitive and specific detection of ligands using engineered riboswitchesen_US
dc.typearticleen_US
dc.contributor.institutionauthorAggrey-Fynn, Joana Efua
dc.relation.journalJournal of Biotechnologyen_US
dc.contributor.departmentIzmir Institute of Technology. Molecular Biology and Geneticsen_US
dc.identifier.volume272-273en_US
dc.identifier.startpage22en_US
dc.identifier.endpage32en_US
dc.identifier.wosWOS:000430278600004
dc.identifier.scopusSCOPUS:2-s2.0-85043536872
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record