Open AccessTriple-strand-forming methylphosphonate oligodeoxynucleotides targeted to mRNA efficiently block protein synthesis.
Author(s) -
Mark A. Reynolds,
Lyle J. Arnold,
M T Almazan,
Terry Beck,
Richard I. Hogrefe,
M D Metzler,
Silva Stoughton,
Ben Tseng,
Tina L. Trapane,
Paul O. P. Ts’o
Publication year - 1994
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.91.26.12433
Subject(s) - chloramphenicol acetyltransferase , oligonucleotide , rna , messenger rna , reverse transcriptase , translation (biology) , triple helix , microbiology and biotechnology , biology , start codon , chemistry , biochemistry , dna , stereochemistry , gene , gene expression , reporter gene
Antisense oligonucleotides are ordinarily targeted to mRNA by double-stranded (Watson-Crick) base recognition but are seldom targeted by triple-stranded recognition. We report that certain all-purine methylphosphonate oligodeoxyribonucleotides (MPOs) form stable triple-stranded complexes with complementary (all-pyrimidine) RNA targets. Modified chloramphenicol acetyltransferase mRNA targets were prepared with complementary all-pyrimidine inserts (18-20 bp) located immediately 3' of the initiation codon. These modified chloramphenicol acetyltransferase mRNAs were used together with internal control (nontarget) mRNAs in a cell-free translation-arrest assay. Our data show that triple-strand-forming MPOs specifically inhibit protein synthesis in a concentration-dependent manner (> 90% at 1 microM). In addition, these MPOs specifically block reverse transcription in the region of their complementary polypyrimidine target sites.