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Title 

Target site search and effective inhibition of leukaemic cell growth by a covalently closed multiple anti-sense oligonucleotide to c-myb

Authors 

Ik Jae MoonYoung Ik LeeChun Sik KwakJe Ho LeeKyu Sam ChoiAlan D SchreiberJong Gu Park

Publisher 

Portland Press

Issue Date 

2000

Citation 

Biochemical Journal, vol. 346, no. 2, pp. 295-303

Keywords 

c-mybgrowth inhibitionoligonucleotide stability

Abstract 

Systematic secondary structure simulation of a target mRNA sequence is shown to be effective for locating a good anti-sense target site. Multiple selected anti-sense sequences were placed in a single molecule. The anti-sense oligonucleotide (oligo) was covalently closed to avoid exonuclease activities and was designated CMAS (covalently closed multiple anti-sense)-oligo. CMAS-oligo was found to be stable, largely preserving its structural integrity after 24 h of incubation in the presence of either exonuclease III or serum. When human c-myb mRNA was targeted by the c-myb CMAS-oligo, expression of the gene was completely abolished. Further, tumour cell growth was inhibited by 82 ± 3% as determined by an MTT [3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay and by 90 ± 1% by [3H]thymidine incorporation. When a leukaemic cell line K562 was treated with CMAS-oligo, colony formation on soft agarose was also decreased by 93%. In contrast, treatment with a scrambled control oligo did not significantly inhibit leukaemic cell growth. These results suggest that a rational target site search is possible for an anti-sense oligo and that CMAS-oligo can be employed as an effective anti-sense agent with enhanced stability.

ISSN 

0264-6021

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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