Title Regioizomerinių SNAP-tag substratų su spiro-izobenzfuran1(3H)-ono fragmentu sintezė ir charakterizavimas
Translation of Title Synthesis and characterization of SNAP-tag substrates containing regioisomeric spiro-isobenzofuran-1(3H)-one fragment.
Authors Markevičius, Vytenis
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Pages 56
Keywords [eng] fluorescent markers ; xanthene analogues ; SNAP-tag substrates
Abstract [eng] All experimental studies were conducted during internships at Max Planck Institute for Biophysical Chemistry, Chromatin Labeling and Imaging research group led by Dr. Gražvydas Lukinavičius. Guanine compounds have been broadly studied for their influence on various cellular processes. Compounds containing this fragment often show cytotoxic and / or protein inhibitory effect. Therefore, they are widely studied and tested for their potential anticancer and antiviral activity. This type of compounds has also potential for the development of fluorescent biomarkers that selectively bind to the SNAP-tag protein. Such biomarkers allow to create new possibilities for the application in fluorescence microscopy visualizing the structure and process of biomolecules or cells. Fluorescent SNAP-tag substrates can be synthesized employing coupling O 6 -substituted guanine derivatives with a fluorophore. Rhodamines or their analogues with high quantum yields are common fluorophores used for construction of such substrates. In this work it was attempted to evaluate the influence of the linker and fluorophore connection position in the marker on its optical properties and ability to penetrate cells. The aim of this work was to synthesize and characterize SNAP-tag substrates using various regioisomers of four fluorophores. During the amide bond formation, fluorophores were coupled with O 6 -[4-(aminomethyl)benzyl]guanine, O 6 -[3-(aminomethyl)benzyl]guanine or 4-{[4- (aminomethyl)benzyl]oxy}-6-chloropyrimidin-2-amine. The optical properties of all goal products were measured and compared. All these substrates were tested in vitro employing fluorescence broadfield microscopy studies with genetically modified U-2 OS H4-SNAP cells.
Dissertation Institution Kauno technologijos universitetas.
Type Master thesis
Language Lithuanian
Publication date 2021