Chemical synthesis of LNA-2-thiouridine and its influence on stability and selectivity of oligonucleotide binding to RNA.
Hybridization to RNA is important for many applications, including antisense therapeutics, RNA interference, and microarray screening. Similar thermodynamic stabilities of A-U and G-U base pairs result in difficulties in selective binding to RNA. Moreover, A-U pairs are weaker than G-C pairs so that binding is sometimes weak when many A-U pairs are present. It is known, however, that replacement of uridine with 2-thiouridine significantly improves binding and selectivity. To test for additional improvement of binding and of the specificity for binding A over G, LNA-2-thiouridine was synthesized for the first time and incorporated into many LNA-2'-O-methyl-RNA/RNA duplexes. UV melting was used to measure the thermodynamic effect of replacing 2'-O-methyluridine with 2'-O-methyl-2-thiouridine or LNA-2-thiouridine. The 2-thiouridine usually enhances binding and selectivity. Selectivity is optimized when a single 2-thiouridine is placed at an internal position in a duplex.
Carlucci
Marta
M
Institute of Bioorganic Chemistry, Polish Academy of Sciences, 61-704 Poznan, Noskowskiego 12/14, Poland.
Kierzek
Elzbieta
E
Olejnik
Anna
A
Turner
Douglas H
DH
Kierzek
Ryszard
R
D009693:Nucleic Acid Hybridization; D009711:Nucleotides / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D011721:Pyrenes / Q000737:chemistry; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry*; D012330:RNA, Double-Stranded / Q000737:chemistry*; D012266:Ribose / Q000031:analogs & derivatives* / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics; D013891:Thiouridine / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry; D044366:Transition Temperature; D014529:Uridine / Q000031:analogs & derivatives / Q000737:chemistry
C434798:2'-O-4'-C-methylene-ribofuranosyl nucleotide; C105273:2-thiouridine; D009711:Nucleotides; D009841:Oligonucleotides; D011721:Pyrenes; D012330:RNA, Double-Stranded; D013891:Thiouridine; D012313:RNA; D012266:Ribose; C030984:pyrene; D014529:Uridine