Project (Gražvydas Lukinavičius)

Methyltransferase-assisted labeling of nucleic acids relies on cofactor S Adenosyl L methionine (AdoMet) analogs where transferrable methyl group has been replaced with extended functionalized chains. However, this approach is largely limited to in vitro applications because of poor cell membrane permeability of AdoMet analogs. The highly interdisciplinary project aims to develop cell membrane permeable AdoMet analogs and use them for DNA or RNA modifying enzymes which can position fluorescent probes at the specific sequences. It involves chemical synthesis of the cofactor analogues and mutagenesis of the enzymes acting on nucleic acids combined with selection of the variants which can incorporate fluorescent labels into natural DNA or RNA. The final goal is to use such labelling strategy in combination with STED or MINFLUX microscopy methods for the elucidation of chromatin structure in living and fixed cells.

• Bucevičius J, Gerasimaitė R, Kiszka KA, Pradhan S, Kostiuk G, Koenen T, Lukinavičius G (2022) A general, highly efficient and facile synthesis of biocompatible rhodamine dyes and probes for live-cell multicolor nanoscopy. bioRxiv 2022.06.13.495683; doi: 10.1101/2022.06.13.495683
  
  
• Gerasimaitė R, Bucevičius J, Kiszka KA, Schnorrenberg S, Kostiuk G, Koenen T, Lukinavičius G (2021) ACS Chem Biol 16 (11), 2130-2136 doi: 10.1021/acschembio.1c00538
  
  
• Bucevičius J, Gilat T, Lukinavičius G (2020) Far-red switching DNA probes for live cell nanoscopy. Chem Commun 56: 14797-14800. doi: 10.1039/D0CC06759H
  
  
• Bucevičius J, Kostiuk G, Gerasimaitė R, Gilat T, Lukinavičius G (2020) Enhancing the biocompatibility of rhodamine fluorescent probes by a neighbouring group effect. Chem Sci 11, 7313-7323. doi: 10.1039/D0SC02154G
  
  
• Bucevičius J, Keller-Findeisen J, Gilat T, Hell SW, Lukinavičius G (2019) Rhodamine-Hoechst positional isomers for highly efficient staining of heterochromatin. Chem Sci 10:1962-1970. doi:10.1039/c8sc05082a
  
  
• 6. Lukinavičius, G., Tomkuvienė, M., Masevičius, V., and Klimašauskas, S. (2013) Enhanced chemical stability of AdoMet analogues for improved methyltransferase-directed labeling of DNA, ACS Chem Biol 8, 1134-1139