Intracellular Expression of a Fluorogenic DNA Aptamer Using Retron Eco2

Author:

Vibhute Mahesh A1ORCID,Machatzke Corbin1,Bigler Katrin1ORCID,Krümpel Saskia1,Summerer Daniel1ORCID,Mutschler Hannes1ORCID

Affiliation:

1. Department of Chemistry and Chemical Biology, TU Dortmund University

Abstract

DNA aptamers are short, single-stranded DNA molecules that bind specifically to a range of targets such as proteins, cells, and small molecules. Typically, they are utilized in the development of therapeutic agents, diagnostics, drug delivery systems, and biosensors. Although aptamers perform well in controlled extracellular environments, their intracellular use has been less explored due to challenges of expressing them in vivo. In this study, we employed the bacterial retron system Eco2, to express a DNA light-up aptamer in Escherichia coli . Both in vitro and in vivo assays confirm that structure-guided insertion of the aptamer domain into the non-coding region of the retron enables reverse transcription and folding of functional aptamer constructs in vivo. Notably, we find only a limited correlation between in vitro and in vivo aptamer performance, suggesting marked folding differences between the two environments. Our findings demonstrate that retrons can be used to effectively express short DNA aptamers within living cells, potentially broadening and optimizing their application in intracellular settings.

Publisher

eLife Sciences Publications, Ltd

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