Year: 2026
Type: preprint
Abstract: Detection of molecules with cell-based sensors allows for conversion of binding events into gene expression outputs. Here, we present a cell-based sensor that can detect extracellular double-stranded DNA. This sensor is based on an engineered receptor which we call Luminescent Ultrasensitive Nucleic Acid Reporter, or LUNAR. LUNAR is based on a recently developed Programmable Antigen-gated G-protein-coupled Engineered Receptor (PAGER). PAGERs are a genetic fusion of an auto-inhibitory peptide, a protein-binding domain, and a modified kappa opioid receptor. PAGERs are gated by two binding events. First, a protein ligand displaces an intramolecular inhibitor, Arodyn, then a second ligand activates the receptor. By replacing the protein-binding domain with a DNA binding zinc finger protein (ZFP) we could detect extracellular DNA in a dose-dependent fashion. Here, we show that first-generation LUNAR constructs can detect both oligonucleotides and plasmid double-stranded DNA with nanomolar sensitivity in mammalian cells. Future work will focus on improving sensitivity, fold-change, and multiplexing capabilities for sequence-specific DNA detection.
Authors: Boao Xia, Nicholas A. Kalogriopoulos, Ruoxin Wen, Zachary M. Lane, Honghao Li, Nicolas Buitrago, Sangsin Lee, Richard D. Gao, Isaac Ive, Yerim Kim, Alice Y. Ting, Jerzy O. Szablowski
Institutions: Rice University, Stanford University
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7203839690
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