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  A Dynamic FRET Reporter of Gene Expression Improved by Functional Screening

Schifferer, M., & Griesbeck, O. (2012). A Dynamic FRET Reporter of Gene Expression Improved by Functional Screening. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 134(37), 15185-15188. doi:10.1021/ja3055673.

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 Creators:
Schifferer, Martina1, Author           
Griesbeck, Oliver1, Author           
Affiliations:
1Research Group: Cellular Dynamics / Griesbeck, MPI of Neurobiology, Max Planck Society, ou_1113560              

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Free keywords: GREEN FLUORESCENT PROTEIN; MESSENGER-RNA; PEPTIDE; BINDING; VISUALIZATION; TRANSCRIPTION; LOCALIZATION; MUTANTS; CELLS
 Abstract: Here, we describe a reporter system that consists of a FRET biosensor and its corresponding aptamer. The FRET biosensor employs the synthetic aptamer binding peptide Rsg1.2 sandwiched between mutants of the Green Fluorescent Protein and undergoes FRET when binding its corresponding Rev Responsive Element (RRE) RNA aptamer. We developed a novel approach to engineer FRET biosensors by linker extension and screening to improve signal strength of the biosensor which we called VAmPIRe (Viral Aptamer binding Peptide based Indicator for RNA detection). We demonstrate that the system is quantitative, reversible and works with high specificity in vitro and in vivo in living bacteria and mammalian cells. Thus, VAmPIRe may become valuable for RNA localizations and as a dynamic RNA-based reporter for live cell imaging. Moreover, functional screening of large libraries as demonstrated here may become applicable to optimize some of the many FRET biosensors of cellular signaling.

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Language(s): eng - English
 Dates: 2012-09-19
 Publication Status: Issued
 Pages: 4
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000308830600006
DOI: 10.1021/ja3055673
 Degree: -

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Title: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Source Genre: Journal
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Publ. Info: 1155 16TH ST, NW, WASHINGTON, DC 20036 USA : AMER CHEMICAL SOC
Pages: - Volume / Issue: 134 (37) Sequence Number: - Start / End Page: 15185 - 15188 Identifier: ISSN: 0002-7863