Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Chemistry, Medicinal, Pharmacology & Pharmacy, cycloaddition, DNA polymerase, fluorescence, imaging agents, oligonucleotides, DIELS-ALDER REACTIONS, TRANS-CYCLOOCTENE, CLICK CHEMISTRY, TETRAZINE, CYCLOADDITIONS, RNA, TRIPHOSPHATES, CONJUGATION, BIOLOGY, Base Sequence, Cyclopropanes, DNA, DNA Primers, Deoxyadenosines, Fluorescent Dyes, HeLa Cells, Humans, Methylation, Microscopy, Fluorescence, Optical Imaging, Hela Cells, 0304 Medicinal and Biomolecular Chemistry, 0601 Biochemistry and Cell Biology, Organic Chemistry, 3101 Biochemistry and cell biology, 3404 Medicinal and biomolecular chemistry

Abstract:

A deoxyadenosine triphosphate (dATP) analogue for DNA labeling was synthesized with the 1-methylcyclopropene (1MCP) group at the 7-position of 7-deaza-2'-deoxyadenosine and applied for primer extension experiments. The real-time kinetic data reveals that this 1MCP-modified dATP analogue is incorporated into DNA much faster than that of the similarly 1MCP-modified deoxyuridine triphosphate (dUTP) analogue. The postsynthetic fluorescent labeling of these oligonucleotides works efficiently according to PAGE analysis, and can be applied for immobilization of a functional antibody on a surface. Site-specific labeling at two different positions in DNA was achieved and the bioorthogonality of the postsynthetic fluorescent labeling was demonstrated in living HeLa cells.