Cross-coupling reactions of nucleoside triphosphates followed by polymerase incorporation. Construction and applications of base-functionalized nucleic acids.
Construction of functionalized nucleic acids (DNA or RNA) via polymerase incorporation of modified nucleoside triphosphates is reviewed and selected applications of the modified nucleic acids are highlighted. The classical multistep approach for the synthesis of modified NTPs by triphosphorylation of modified nucleosides is compared to the novel approach consisting of direct aqueous cross-coupling reactions of unprotected halogenated nucleoside triphosphates. The combination of cross-coupling of NTPs with polymerase incorporation gives an efficient and straightforward two-step synthesis of modified nucleic acids. Primer extension using biotinylated templates followed by separation using streptavidine-coated magnetic beads and DNA duplex denaturation is used for preparation of modified single stranded oligonucleotides. Examples of using this approach for electrochemical DNA labelling and bioanalytical applications are given.
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, CZ-16610 Prague 6, Czech Republic. hocek@uochb.cas.cz
Michal
M
Hocek
Miroslav
M
Fojta
D001483:Base Sequence; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D009696:Nucleic Acids / Q000378:metabolism*; D009711:Nucleotides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011122:Polyphosphates / Q000737:chemistry*
D009696:Nucleic Acids; D009711:Nucleotides; D011122:Polyphosphates; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase; C005692:triphosphoric acid