biostudies-literatureXiong QFundamental Research Funds for the Central UniversitiesNational Natural Science Foundation of ChinaSichuan University3571-3581https://www.ebi.ac.uk/biostudies/studies/S-EPMC8943893biostudies-literatureUnknown13(12)Genetic incorporation of novel noncanonical amino acids (ncAAs) that are specialized for the photo-click reaction allows the precisely orthogonal and site-specific functionalization of proteins in living cells under photo-control. However, the development of a r̲ing-strain <i>i̲n situ</i> l̲oadable d̲ipolarophile (RILD) as a genetically encodable reporter for photo-click bioconjugation with spatiotemporal controllability is quite rare. Herein, we report the design and synthesis of a photo-switchable d̲ib̲enzo[<i>b</i>,<i>f</i>][1,4,5]t̲hiad̲iazepine-based a̲lanine (DBTDA) ncAA, together with the directed evolution of a pyrrolysyl-tRNA synthetase/tRNA_CUA pair (PylRS/tRNA_CUA), to encode the DBTDA into recombinant proteins as a RILD in living <i>E. coli</i> cells. The faChemical scienceExpanding the functionality of proteins with genetically encoded dibenzo[<i>b</i>,<i>f</i>][1,4,5]thiadiazepine: a photo-transducer for photo-click decoration.PMC89438932020SCUNL1052207709020826041D411722001181Wang CXiong QFu JLi BYu ZShen XZheng TZhao XfalseExpanding the functionality of proteins with genetically encoded dibenzo[<i>b</i>,<i>f</i>][1,4,5]thiadiazepine: a photo-transducer for photo-click decoration.Genetic incorporation of novel noncanonical amino acids (ncAAs) that are specialized for the photo-click reaction allows the precisely orthogonal and site-specific functionalization of proteins in living cells under photo-control. However, the development of a r̲ing-strain <i>i̲n situ</i> l̲oadable d̲ipolarophile (RILD) as a genetically encodable reporter for photo-click bioconjugation with spatiotemporal controllability is quite rare. Herein, we report the design and synthesis of a photo-switchable d̲ib̲enzo[<i>b</i>,<i>f</i>][1,4,5]t̲hiad̲iazepine-based a̲lanine (DBTDA) ncAA, together with the directed evolution of a pyrrolysyl-tRNA synthetase/tRNA_CUA pair (PylRS/tRNA_CUA), to encode the DBTDA into recombinant proteins as a RILD in living <i>E. coli</i> cells. The fa2022-01-01T00:00:00Z2022 Mar2026-05-09T15:10:36.12Z2024-11-05T17:03:08.591ZS-EPMC89438933543285610.1039/d1sc05710c