Project (Gražvydas Lukinavičius)

Covalent probes (Cops) for chromatin nanoscopy

Development of super-resolution microscopy (nanoscopy) has provided an opportunity for monitoring the cellular structures with an unprecedented resolution in living cells. Traditionally, proteins of interest (POI’s) in living cells are imaged by expressing them as fusions with fluorescent proteins (FP’s). However, FP have certain limitations: relatively large size of FP and possibility to attach only to the C- or N-terminus of the POI in some cases interfere with the cellular functions of POI; their photostability is sometimes inferior to that of small molecule fluorophores; the genetic modification required largely limits the use of FP’s in patient-derived cells for the diagnostics. Therefore in certain cases the small molecule probes might clearly be advantageous and possess large potential for the imaging applications. A very attractive approach would be to design a POI-specific probe which is able to form covalent bond and does not require any genetic manipulations. For example, such probe could consist of a fluorophore, a chemically active linker and a targeting moiety. Ligand binding guides the probe to POI where a cleavable linker can react with a nucleophile located near the binding site. After the reaction targeting moiety can dissociate and only fluorophore remains attached to POI. The project aims to create such probes for the chromatin components.

Homepage Research Group

For more information see for instance:

  • Kostiuk G, Bucevičius J, Gerasimaitė R, Lukinavičius G (2019) Application of STED imaging for chromatin studies. Journal of Physics D: Applied Physics 52:504003. doi:10.1088/1361-6463/ab4410

  • Bucevicius 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

  • Lukinavičius G, Reymond L, Umezawa K, Sallin O, D'Este E, Gottfert F, Ta H, Hell SW, Urano Y, Johnsson K (2016) Fluorogenic Probes for Multicolor Imaging in Living Cells. J Am Chem Soc 138:9365-9368. doi:10.1021/jacs.6b04782

  • Lukinavičius G, Blaukopf C, Pershagen E, Schena A, Reymond L, Derivery E, Gonzalez-Gaitan M, D'Este E, Hell SW, Gerlich DW, Johnsson K (2015) SiR-Hoechst is a far-red DNA stain for live-cell nanoscopy. Nat Commun 6:8497. doi:10.1038/ncomms9497