Programmable eukaryotic protein synthesis with RNA sensors by harnessing ADAR.

Autor: Jiang K; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA., Koob J; Broad Institute of MIT and Harvard, Cambridge, MA, USA.; Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.; Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA., Chen XD; Broad Institute of MIT and Harvard, Cambridge, MA, USA.; Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.; Systems, Synthetic, and Quantitative Biology Program, Harvard Medical School, Boston, MA, USA., Krajeski RN; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA., Zhang Y; Broad Institute of MIT and Harvard, Cambridge, MA, USA., Volf V; Broad Institute of MIT and Harvard, Cambridge, MA, USA.; Department of Genetics, Harvard Medical School, Boston, MA, USA., Zhou W; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA., Sgrizzi SR; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA., Villiger L; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA., Gootenberg JS; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA. jgoot@mit.edu.; Gene Regulation Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA. jgoot@mit.edu., Chen F; Broad Institute of MIT and Harvard, Cambridge, MA, USA. chenf@broadinstitute.org.; Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA. chenf@broadinstitute.org.; Gene Regulation Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA. chenf@broadinstitute.org., Abudayyeh OO; McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA. omar@abudayyeh.science.; Gene Regulation Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA. omar@abudayyeh.science.
Jazyk: angličtina
Zdroj: Nature biotechnology [Nat Biotechnol] 2023 May; Vol. 41 (5), pp. 698-707. Date of Electronic Publication: 2022 Oct 27.
DOI: 10.1038/s41587-022-01534-5
Abstrakt: Programmable approaches to sense and respond to the presence of specific RNAs in biological systems 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 protein by the presence of endogenous RNA transcripts. Introduction of a stop codon in a guide upstream of the cargo makes translation contingent on binding of an endogenous transcript to the guide, leading to ADAR editing of the stop codon and allowing translational readthrough. Through systematic sensor engineering, we achieve 277 fold improvement in sensor activation and engineer RADARS with diverse cargo proteins, including luciferases, fluorescent proteins, recombinases, and caspases, enabling 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.
(© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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