{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:45:33Z","timestamp":1768506333329,"version":"3.49.0"},"posted":{"date-parts":[[2023,7,30]]},"group-title":"Biochemistry","reference-count":63,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,12,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Fluorescent light-up aptamer (FLAP) systems are promising biosensing platforms that can be genetically encoded. Here, we describe how a single FLAP that works with specific organic ligands can detect multiple, structurally unique, non-fluorogenic, and reactive inorganic targets. We developed 4-\n                  <jats:italic>O<\/jats:italic>\n                  -functionalized benzylidene imidazolinones as pre-ligands with suppressed fluorescent binding interactions with the RNA aptamer Baby Spinach. Inorganic targets, hydrogen sulfide (H\n                  <jats:sub>2<\/jats:sub>\n                  S) or hydrogen peroxide (H\n                  <jats:sub>2<\/jats:sub>\n                  O\n                  <jats:sub>2<\/jats:sub>\n                  ), can specifically convert these pre-ligands into the native benzylidene imidazolinones, and thus be detected with Baby Spinach. Adaptation of this approach to live cells opened a new opportunity for top-down construction of whole-cell sensors:\n                  <jats:italic>Escherichia coli<\/jats:italic>\n                  transformed with a Baby Spinach-encoding plasmid and incubated with pre-ligands generated fluorescence in response to exogenous H\n                  <jats:sub>2<\/jats:sub>\n                  S or H\n                  <jats:sub>2<\/jats:sub>\n                  O\n                  <jats:sub>2<\/jats:sub>\n                  . Our approach eliminates the requirement of in vitro selection of a new aptamer sequence for molecular target detection, allows for the detection of short-lived targets, thereby advancing FLAP systems beyond their current capabilities. Leveraging the functional group reactivity of small molecules can lead to cell-based sensors for inorganic molecular targets, exploiting a new synergism between synthetic organic chemistry and synthetic biology.\n                <\/jats:p>","DOI":"10.1101\/2023.07.29.551132","type":"posted-content","created":{"date-parts":[[2023,7,31]],"date-time":"2023-07-31T00:10:10Z","timestamp":1690762210000},"source":"Crossref","is-referenced-by-count":1,"title":["A Small-Molecule Approach to Bypass In Vitro Selection of New Aptamers: Designer Pre-Ligands Turn Baby Spinach into Sensors for Reactive Inorganic 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