Title:
Dual FRET molecular beacons for mRNA detection in living cells

dc.contributor.author Santangelo, Philip J. en_US
dc.contributor.author Nix, Brent en_US
dc.contributor.author Tsourkas, Andrew en_US
dc.contributor.author Bao, Gang en_US
dc.contributor.corporatename Georgia Institute of Technology. Dept. of Biomedical Engineering en_US
dc.contributor.corporatename Emory University. Dept. of Biomedical Engineering en_US
dc.date.accessioned 2013-06-03T20:16:36Z
dc.date.available 2013-06-03T20:16:36Z
dc.date.issued 2004
dc.description © The Author(s) 2004. Published by Oxford University Press.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. en_US
dc.description DOI: 10.1093/nar/gnh062 en_US
dc.description.abstract The ability to visualize in real-time the expression level and localization of specific endogenous RNAs in living cells can offer tremendous opportunities for biological and disease studies. Here we demonstrate such a capability using a pair of molecular beacons, one with a donor and the other with an acceptor fluorophore that hybridize to adjacent regions on the same mRNA target, resulting in fluorescence resonance energy transfer (FRET). Detection of the FRET signal significantly reduced false positives, leading to sensitive imaging of K-ras and survivin mRNAs in live HDF and MIAPaCa-2 cells. FRET detection gave a ratio of 2.25 of K-ras mRNA expression in stimulated and unstimulated HDF, comparable to the ratio of 1.95 using RT±PCR, and in contrast to the single-beacon result of 1.2. We further revealed intriguing details of K-ras and survivin mRNA localization in living cells. The dual FRET molecular beacons approach provides a novel technique for sensitive RNA detection and quantification in living cells. en_US
dc.identifier.citation Philip J. Santangelo, Brent Nix, Andrew Tsourkas and Gang Bao, “Dual FRET molecular beacons for mRNA detection in living cells,” Nucleic Acids Research, 32, e57, 2004. en_US
dc.identifier.doi 10.1093/nar/gnh062
dc.identifier.issn 0305-1048
dc.identifier.uri http://hdl.handle.net/1853/47167
dc.language.iso en_US en_US
dc.publisher Georgia Institute of Technology en_US
dc.publisher.original Oxford University Press en_US
dc.subject Real-time visualization en_US
dc.subject RNA en_US
dc.subject Fluorescence resonance energy transfer en_US
dc.subject FRET en_US
dc.subject Molecular beacons en_US
dc.title Dual FRET molecular beacons for mRNA detection in living cells en_US
dc.type Text
dc.type.genre Article
dspace.entity.type Publication
local.contributor.corporatename Wallace H. Coulter Department of Biomedical Engineering
local.contributor.corporatename College of Engineering
relation.isOrgUnitOfPublication da59be3c-3d0a-41da-91b9-ebe2ecc83b66
relation.isOrgUnitOfPublication 7c022d60-21d5-497c-b552-95e489a06569
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
bme_Bao_003.pdf
Size:
334.05 KB
Format:
Adobe Portable Document Format
Description: