Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus.

Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus. Research Abstract Details 

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Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus. Abstract Text:

Mariana Kiomy Osako,Naruya Tomita,Hironori Nakagami,Yasuo Kunugiza,Masafumi Yoshino,Kazuhiko Yuyama,Tetsuya Tomita,Hideki Yoshikawa,Toshio Ogihara,Ryuichi Morishita,Mariana Kiomy Osako,Naruya Tomita,Hironori Nakagami,Yasuo Kunugiza,Masafumi Yoshino,Kazuhiko Yuyama,Tetsuya Tomita,Hideki Yoshikawa,Toshio Ogihara,Ryuichi Morishita,

BACKGROUND: For the development of molecular therapy based on oligodeoxynucleotides (ODN), ODN have to be stable against nucleases and be specific to the target transcription factor. To decrease non-specific binding and degradation from the 3'-terminus of ODN, we designed partially annealed ODN by binding the extremities of two single strands, resulting in a ribbon-shaped ODN, so called ribbon-type decoy ODN (R-ODN). METHODS: We evaluated the efficiency in the process of enzymatic ligation of R-ODN, the binding activity to nuclear factor-kappaB (NF-kappaB), and the stability against Exonuclease III and nucleases present in serum. The functional activity of R-ODN to inhibit NF-kappaB in vitro was evaluated in human aortic smooth muscle cells (VSMC): TNF-alpha-induced proliferation rate and MMP-9 expression were assessed after R-ODN transfection. RESULTS AND CONCLUSIONS: Although R-ODN have a phosphodiester backbone, their physical conformation was designed to provide nuclease resistance without interfering with their binding activity. As expected, R-ODN showed more resistance to exonucleases and stability in 100% serum than non-modified decoy ODN (N-ODN). Importantly, the R-ODN construction did not interfere with its binding activity to NF-kappaB, similar to N-ODN. Transfection of R-ODN significantly inhibited the expression of MMP-9 induced by TNF-alpha in VSMC as assessed by real-time polymerase chain reaction (PCR), and R-ODN also inhibited the proliferation of VSMC induced by TNF-alpha (10 ng/ml), similar to phosphorothioate decoy ODN. Overall, the development of ribbon NF-kappaB decoy ODN could provide a useful tool for basic and clinical research.

Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus. Publishing Authors By Initials

MK Osako,N Tomita,H Nakagami,Y Kunugiza,M Yoshino,K Yuyama,T Tomita,H Yoshikawa,T Ogihara,R Morishita,MK Osako,N Tomita,H Nakagami,Y Kunugiza,M Yoshino,K Yuyama,T Tomita,H Yoshikawa,T Ogihara,R Morishita,

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Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: The journal of gene medicine

VOLUME: 9

Page Numbers: 812-9

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ISSN: 1099-498X

DAY: 27

MONTH: Sep

YEAR: 2007

Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus. Information

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LANGUAGE: eng

NlmUniqueID: 9815764

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Grant and Affiliation Information for Increase in nuclease resistance and incorporation of NF-kappaB decoy oligodeoxynucleotides by modification of the 3'-terminus.

AFFILIATION: Division of Clinical Gene Therapy, Osaka University Graduate School of Medicine, 2-2 Yamada-Oka, Suita, Osaka, Japan.

Country: England

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MEDLINETA: J Gene Med

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