Fiche publication
Date publication
octobre 2009
Journal
Nucleic acids research
Auteurs
Membres identifiés du Cancéropôle Est :
Dr REMY Jean-Serge
Tous les auteurs :
Moreau V, Voirin E, Paris C, Kotera M, Nothisen M, Rémy JS, Behr JP, Erbacher P, Lenne-Samuel N
Lien Pubmed
Résumé
Most nucleic acid-based technologies rely upon sequence recognition between an oligonucleotide and its nucleic acid target. With the aim of improving hybridization by decreasing electrostatic repulsions between the negatively charged strands, novel modified oligonucleotides named Zip nucleic acids (ZNAs) were recently developed. ZNAs are oligonucleotide-oligocation conjugates whose global charge is modulated by the number of cationic spermine moieties grafted on the oligonucleotide. It was demonstrated that the melting temperature of a hybridized ZNA is easily predictable and increases linearly with the length of the oligocation. Furthermore, ZNAs retain the ability to discriminate between a perfect match and a single base-pair-mismatched complementary sequence. Using quantitative PCR, we show here that ZNAs are specific and efficient primers displaying an outstanding affinity toward their genomic target. ZNAs are particularly efficient at low magnesium concentration, low primer concentrations and high annealing temperatures, allowing to improve the amplification in AT-rich sequences and potentially multiplex PCR applications. In reverse transcription experiments, ZNA gene-specific primers improve the yield of cDNA synthesis, thus increasing the accuracy of detection, especially for genes expressed at low levels. Our data suggest that ZNAs exhibit faster binding kinetics than standard and locked nucleic acid-containing primers, which could explain why their target recognition is better for rare targets.
Mots clés
AT Rich Sequence, DNA Primers, Polymerase Chain Reaction, standards, Reverse Transcription
Référence
Nucleic Acids Res.. 2009 Oct;37(19):e130
