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bioRxiv · 10.64898/2026.08.18.745542

Glycine detection with a nuclease-stable L-RNA sensor

Bodin, M. R.·Han, X.·Sczepanski, J. T.·Hammond, M. C.
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Abstract

Glycine is a vital extracellular signal in bacteria, plants, and the brain. Although RNA-based sensors detect glycine in cells, their extracellular application in native biological environments is limited by enzymatic degradation from nucleases. Mirror-image RNA is nuclease-resistant and preserves the tertiary structure required for RNA function, but synthesizing long L-RNAs such as the 170-nt glycine sensor (glyS) remains challenging. Here, we applied cross-chiral ligation with natural D-RNA ribozymes to assemble a mirror-image L-RNA glycine sensor (L-glyS). Optimization of the ligation conditions enabled up to 68% conversion to the full-length sensor. L-glyS displayed nuclease resistance and maintained glycine-dependent fluorescence in serum, where the original D-glyS lost function. These results establish cross-chiral ligation as a strategy for constructing long, functional L-RNAs and broaden the possible applications of RNA-based sensors to extracellular detection of small molecules.

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Bodin, M. R., Han, X., Sczepanski, J. T., Hammond, M. C.. 2026-08-21. Glycine detection with a nuclease-stable L-RNA sensor. https://doi.org/10.64898/2026.08.18.745542

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