Triple-strand-forming methylphosphonate oligodeoxynucleotides targeted to mRNA efficiently block protein synthesis.
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.
Reynolds
M A
MA
Genta, Inc., San Diego, CA 92121.
Arnold
L J
LJ
Almazan
M T
MT
Beck
T A
TA
Hogrefe
R I
RI
Metzler
M D
MD
Stoughton
S R
SR
Tseng
B Y
BY
Trapane
T L
TL
Ts'o
P O
PO
D001483:Base Sequence; D002474:Cell-Free System; D017931:DNA Primers / Q000737:chemistry; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D063065:Organophosphonates; D014176:Protein Biosynthesis; D011687:Purines / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry*; D018894:Reverse Transcriptase Inhibitors; D013329:Structure-Activity Relationship
D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D063065:Organophosphonates; D011687:Purines; D012333:RNA, Messenger; D018894:Reverse Transcriptase Inhibitors