Intracellular Expression of a Fluorogenic DNA Aptamer Using Retron Eco2
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 inEscherichia 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.
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