Targeting RNA for degradation with a (2'-5')oligoadenylate-antisense chimera

• Published in: Proceedings of the National Academy of Sciences - PNAS 1993-02, Vol.90 (4), p.1300-1304
• Main Authors: TORRENCE, P. F, MAITRA, R. K, LESIAK, K, SHAHRZAD KHAMNEI, AIMIN ZHOU, SILVERMAN, R. H
• Format: Article
• Language: English
• Subjects: Adenine Nucleotides; AIDS/HIV; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Bacteriophage T4 - genetics; Base Sequence; Biochemistry; Biological and medical sciences; Chimera; Genes, vif; HIV-1 - genetics; human immunodeficiency virus 1; Humans; Medical sciences; Molecular Sequence Data; Oligonucleotides, Antisense; Oligoribonucleotides; Pharmacology. Drug treatments; Plasmids; Ribonucleases - metabolism; Ribonucleic acid; RNA; RNA - chemistry; RNA - metabolism; RNA, Viral - chemistry; RNA, Viral - metabolism; Transcriptional Activation; Tumor Cells, Cultured
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.4.1300

Antisense oligonucleotides hold considerable promise both as research tools for inhibiting gene expression and as agents for the treatment of a myriad of human diseases. However, targeted destruction of RNA has been difficult to achieve in a versatile, efficient, and reliable manner. We have developed an effective strategy for cleaving unique RNA sequences with 2-5A-dependent RNase, an endoribonuclease that mediates inhibitory effects of interferon on virus infection and is activated by 5'-phosphorylated 2'-5'-linked oligoadenylates known as 2-5A [pn5' A2'(p5' A2')mp5'A], resulting in the cleavage of single-stranded RNA predominantly after UpUp and UpAp sequences. To direct 2-5A-dependent RNase to cleave unique RNA sequences, p5' A2' p5' A2'p5'A was covalently linked to an antisense oligonucleotide to yield a chimeric molecule (2-5A:AS). The antisense oligonucleotide component of 2-5A:AS bound a specific RNA sequence while the accompanying 2-5A component activated 2-5A-dependent RNase, thereby causing the cleavage of the RNA in the targeted sequence. This strategy was demonstrated by inducing specific cleavage within a modified human immunodeficiency virus type 1 vif mRNA in a cell-free system from human lymphoblastoid cells. Because 2-5A-dependent RNase is present in most mammalian cells, the control of gene expression based on this technology--including therapies for cancer, viral infections, and certain genetic diseases--can be envisioned.