Direct enzymatic synthesis of aldehyde-functionalized DNA and its conjugation with hydrazines and amines

0361574 - ÚOCHB 2012 CZ eng C - Conference Paper (international conference)
Raindlová, Veronika - Hocek, Michal
Direct enzymatic synthesis of aldehyde-functionalized DNA and its conjugation with hydrazines and amines.
Chemistry of Nucleic Acid Components. 15th Symposium. Praha: Institute of Organic Chemistry and Biochemistry AS CR, v. v. i., 2011 - (Hocek, M.), s. 441-443. Collection Symposium Series, 12. ISBN 978-80-86241-37-1.
[Chemistry of Nucleic Acid Components /15./. Český Krumlov (CZ), 05.06.2011-10.06.2011]
R&D Projects: GA MŠMT LC512; GA ČR GA203/09/0317
Institutional research plan: CEZ:AV0Z40550506
Keywords : bioconjugation * DNA * aldehydes
Subject RIV: CC - Organic Chemistry

A new simple methodology for DNA conjugation or staining was developed. 2′-Deoxyribonucleoside triphosphates (dNTPs) bearing reactive aldehyde group were prepared by one-step Suzuki cross-coupling reaction of halogenated dNTPs with boronic acid. These modified dNTPs were enzymatically incorporated into DNA by primer extension (PEX) or amplified by polymerase chain reaction (PCR) using different DNA polymerases. The followup reaction between aldehyde-modified PCR products and hydrazine derivatives gave coloured DNA conjugated hydrazones. This methodology was also used for further conjugations of aldehyde-modified 2′-deoxyribonucleoside monophosphates (dNMPs) with amines by reductive amination.
Permanent Link: http://hdl.handle.net/11104/0198862