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Specific inhibition of influenza virus RNA polymerase and nucleoprotein gene expression by circular dumbbell RNA/DNA chimeric oligonucleotides containing antisense phosphodiester oligonucleotides
Author(s) -
Abe Takayuki,
Takai Kazuyuki,
Nakada Susumu,
Yokota Tomoyuki,
Takaku Hiroshi
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00206-3
Subject(s) - oligonucleotide , rnase h , rna , microbiology and biotechnology , phosphodiester bond , dna , biology , sense (electronics) , rnase p , rna dependent rna polymerase , chemistry , gene , biochemistry
We have designed a new class of oligonucleotides, `dumbbell RNA/DNA chimeric oligonucleotides', consisting of a sense RNA sequence and its complementary antisense DNA sequence, with two hairpin loop structures. The reaction of the nicked (NDRDON) and circular (CDRDON) dumbbell RNA/DNA chimeric oligonucleotides with RNase H gave the corresponding antisense phosphodiester oligodeoxynucleotide together with the sense RNA cleavage products. The liberated antisense phosphodiester oligodeoxynucleotide was bound to the target RNA, which gave RNA cleavage products by treatment with RNase H. The circular dumbbell RNA/DNA chimeric oligonucleotide showed more nuclease resistance than the linear antisense phosphodiester oligonucleotide (anti‐ODN) and the nicked dumbbell RNA/DNA chimeric oligonucleotide. The CDRDON with four target sites (influenza virus A RNA polymerases (PB1, PB2, PA) and nucleoprotein (NP)) was synthesized and tested for inhibitory effects by a CAT‐ELISA assay using the clone 76 cell line. The circular dumbbell DNA/RNA chimeric oligonucleotide (CDRDON‐PB2‐as) containing an AUG initiation codon sequence as the target of PB2 showed highly inhibitory effects.