Novel method for covalent fluorescent labeling of plasmid DNA that maintains structural integrity of the plasmid

Carole Neves, Gerardo Byk, Virginie Escriou, Florence Bussone, Daniel Scherman, Pierre Wils

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

We have developed a chemical strategy for covalent coupling of fluorophores to plasmid DNA. A p-azido-tetrafluoro-benzyl-lissamine conjugate was synthesized and purified. This conjugate was used to covalently associate fluorescent molecules to plasmid DNA by photoactivation. In contrast to nick-translated plasmid DNA, plasmid-lissamine conjugates appeared on gel as supercoiled DNA. Reporter gene was expressed after transfection of the plasmid-lissamine conjugates in NIH 3T3 cells, although gene transfer efficiency was decreased by 60% as compared with unlabeled DNA. Intracellular traffic of plasmid-lissamine conjugates was studied in transfected cells. After cytoplasmic microinjection, fluorescent plasmid did not diffuse from the site of injection and appeared to be progressively degraded in the cytoplasm.

Original languageEnglish
Pages (from-to)51-55
Number of pages5
JournalBioconjugate Chemistry
Volume11
Issue number1
DOIs
StatePublished - 2000

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