Biotinylated antisense methylphosphonate oligodeoxynucleotides. Inhibition of spliceosome assembly and affinity selection of U1 and U2 small nuclear RNPs.
Methylphosphonate (PC) backbone oligodeoxynucleotides complementary to the 5'-terminal nucleotides of U1 and U2 small nuclear (sn) RNAs do not elicit RNase H action under conditions in which natural (phosphodiester) oligodeoxynucleotides yield extensive RNase H cleavage. We show here that antisense PC oligonucleotides can mask sites in U1 and U2 snRNPs that are required for spliceosome formation. We further report that biotinylated derivatives of antisense PC oligos can be used for affinity selection of U1 and U2 snRNPs.
Temsamani
J
J
Cell Biology Group, The Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.
Agrawal
S
S
Pederson
T
T
D001483:Base Sequence; D001710:Biotin; D002467:Cell Nucleus / Q000378:metabolism; D004722:Endoribonucleases / Q000378:metabolism; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000494:pharmacology*; D009943:Organophosphorus Compounds; D012326:RNA Splicing / Q000187:drug effects*; D012334:RNA, Neoplasm / Q000187:drug effects / Q000378:metabolism; D016914:Ribonuclease H; D012261:Ribonucleoproteins / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear
D016376:Oligonucleotides, Antisense; D009943:Organophosphorus Compounds; D012334:RNA, Neoplasm; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; C032627:methylphosphonic acid; D001710:Biotin; D004722:Endoribonucleases; D016914:Ribonuclease H