Transcriptome-Wide off-target RNA sequencing for RADARS. Homo sapiens

Programmable approaches to sense and respond to the presence of specific RNAs in biological systems would have broad applications in research, diagnostics, and therapeutics. Here, we engineer a programmable RNA-sensing technology, reprogrammable ADAR sensors (RADARS), which harnesses RNA editing by adenosine deaminases acting on RNA (ADAR) to gate translation of a cargo target protein by the presence of endogenous RNA transcripts. By including a stop codon in a guide upstream of the cargo, translation only occurs when an endogenous transcript is In mammalian cells, we engineer RADARS for diverse target proteins, including luciferases, fluorescent proteins, recombinases, and caspases, achieving detection sensitivity on endogenous transcripts expressed at levels as low as 13 transcripts per million. We show that RADARS are functional as either expressed DNA or synthetic mRNA and with either exogenous or endogenous ADAR. We apply RADARS in multiple contexts, including tracking transcriptional states, RNA-sensing induced cell death, cell type identification, and control of synthetic mRNA translation.