Ma. Reynolds et al., TRIPLE-STRAND-FORMING METHYLPHOSPHONATE OLIGODEOXYNUCLEOTIDES TARGETED TO MESSENGER-RNA EFFICIENTLY BLOCK PROTEIN-SYNTHESIS, Proceedings of the National Academy of Sciences of the United Statesof America, 91(26), 1994, pp. 12433-12437
Antisense oligonucleotides are ordinarily targeted to mRNA by double-s
tranded (Watson-Crick) base recognition but are seldom targeted by tri
ple-stranded recognition. We report that certain all-purine methylphos
phonate oligodeoxyribonucleotides (MPOs) form stable triple-stranded c
omplexes with complementary (all-pyrimidine) RNA targets. Modified chl
oramphenicol acetyltransferase mRNA targets were prepared with complem
entary all-pyrimidine inserts (18-20 bp) located immediately 3' of the
initiation codon. These modified chloramphenicol acetyltransferase mR
NAs were used together with internal control (nontarget) mRNAs in a ce
ll-free translation-arrest assay. Our data show that triple-strand-for
ming MPOs specifically inhibit protein synthesis in a concentration-de
pendent manner (> 90% at 1 mu M). In addition, these MPOs specifically
block reverse transcription in the region of their complementary poly
pyrimidine target sites.