Specific inhibition of influenza virus RNA polymerase and nucleoprotein gene expression by circular dumbbell RNA/DNA chimeric oligonucleotides containing antisense phosphodiester oligonucleotides

FEBS Letters

Nakada, Susumu²  Abe, Takayuki³  Takai, Kazuyuki³  Takaku, Hiroshi³  Yokota, Tomoyuki¹ 

[1] Rational Drug Design Laboratories (R.D.L.), Misato, Matsukawa, Fukushima 960-12, Japan;Institute for Drug Discovery Research, Yamanouchi Pharmaceutical Co., Ltd., Miyukigaoka, Tsukuba, Ibaraki 305, Japan;Department of Industrial Chemistry, Chiba Institute of Technology, Tsudanuma, Narashino, Chiba 275, Japan

关键词: Circular dumbbell RNA/DNA chimeric oligonucleotide;    Antisense phosphodiester oligonucleotide;    Melting temperature;    RNase H;    Exonuclease resistance;    Influenza virus;    RNA polymerase;    Nucleoprotein;    CAT assay;   

DOI  :  10.1016/S0014-5793(98)00206-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown