2'-O-methyl-modified phosphorothioate antisense oligonucleotides have reduced non-specific effects in vitro.

Abstract:

:Antisense oligodeoxynucleotides (ODNs) have biological activity in treating various forms of cancer. The antisense effects of two types of 20mer ODNs, phosphorothioate-modified ODNs (S-ODNs) and S-ODNs with 12 2'-O-methyl groups (Me-S-ODNs), targeted to sites 109 and 277 of bcl-2 mRNA, were compared. Both types were at least as effective as G3139 (Genta, Inc.) in reducing the level of Bcl-2 protein in T24 cells following a 4 h transfection at a dose of 0.1 micro M. Circular dichroism spectra showed that both types formed A-form duplexes with the complementary RNA, and the melting temperatures were in the order of Me-S-ODN.RNA > normal DNA.RNA > S-ODN.RNA. In comparison with the S-ODN, the Me-S-ODN had reduced toxic growth inhibitory effects, was less prone to bind the DNA-binding domain A of human replication protein A, and was as resistant to serum nucleases. Neither type of oligomer induced apoptosis, according to a PARP-cleavage assay. Hybrids formed with Me-S-ODN sequences were less sensitive to RNase H degradation than those formed with S-ODN sequences. Despite this latter disadvantage, the addition of 2'-O-methyl groups to a phosphorothioate-modified ODN is advantageous because of increased stability of binding and reduced non-specific effects.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Yoo BH,Bochkareva E,Bochkarev A,Mou TC,Gray DM

doi

10.1093/nar/gkh516

keywords:

subject

Has Abstract

pub_date

2004-04-02 00:00:00

pages

2008-16

issue

6

eissn

0305-1048

issn

1362-4962

pii

32/6/2008

journal_volume

32

pub_type

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