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In vitro and in vivo targeted delivery of IL-10 interfering RNA by JC virus-like particles.

Chou MI, Hsieh YF, Wang M, Chang JT, Chang D, Zouali M, Tsay GJ - J. Biomed. Sci. (2010)

Bottom Line: In RAW 264.7 cells, IL-10 shRNA was found to reduce IL-10 expression by 85 to 89%, as compared with VLPs alone.IL-10 shRNA did not cross-react with TNF-alpha mRNA or influence the expression of TNF-alpha.In BALB/c mice IL-10 shRNA could reduce 95% of IL-10 secretion.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.

ABSTRACT

Background: RNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Delivering sequences of RNAi in vivo remains a problem. The aim of this study was to use JC virus (JCV) virus-like particles (VLPs) as a vector for delivering RNAi in silencing the cytokine gene of IL-10.

Methods: JCV VLPs were generated by recombinant JCV VP1 protein in yeast expression system. DNA fragment containing IL-10 shRNA was packaged into VLPs by osmotic shock.

Results: In RAW 264.7 cells, IL-10 shRNA was found to reduce IL-10 expression by 85 to 89%, as compared with VLPs alone. IL-10 shRNA did not cross-react with TNF-alpha mRNA or influence the expression of TNF-alpha. In BALB/c mice IL-10 shRNA could reduce 95% of IL-10 secretion. Surprisingly, it also down regulated TNF-alpha expression.

Conclusions: We show for the first time that JCV VLPs empty capsids are competent vectors to deliver RNAi and are nontoxic to cells, suggesting that JCV VLPs is an efficient agent to deliver RNAi in both murine macrophage cells and BALB/c mice. This system provides an efficient means for delivering the RNAi for gene therapy purposes.

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Related in: MedlinePlus

Outcome of BALB/c mice treated with LPS, irrelevant shRNA and IL-10 shRNA. Groups of BALB/c mice were intraperitoneally injected with LPS (25 μg). After two hours, mice received intravenously VLPs-IL-10 shRNA and irrelevant shRNA. The survival rate and survival time of the animals were observed and compared between groups.
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Figure 5: Outcome of BALB/c mice treated with LPS, irrelevant shRNA and IL-10 shRNA. Groups of BALB/c mice were intraperitoneally injected with LPS (25 μg). After two hours, mice received intravenously VLPs-IL-10 shRNA and irrelevant shRNA. The survival rate and survival time of the animals were observed and compared between groups.

Mentions: All mice treated with PBS only or LPS-VLPs-IL-10 shRNA survived in the end of the experiment, but all mice treated with LPS only or LPS-VLPs irrelevant shRNA died after 24 h of treatment. The mice treated with LPS-VLPs-IL-10 shRNA had longer lifespan than those treated with LPS-VLPs irrelevant shRNA or LPS. (Fig. 5)


In vitro and in vivo targeted delivery of IL-10 interfering RNA by JC virus-like particles.

Chou MI, Hsieh YF, Wang M, Chang JT, Chang D, Zouali M, Tsay GJ - J. Biomed. Sci. (2010)

Outcome of BALB/c mice treated with LPS, irrelevant shRNA and IL-10 shRNA. Groups of BALB/c mice were intraperitoneally injected with LPS (25 μg). After two hours, mice received intravenously VLPs-IL-10 shRNA and irrelevant shRNA. The survival rate and survival time of the animals were observed and compared between groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2902427&req=5

Figure 5: Outcome of BALB/c mice treated with LPS, irrelevant shRNA and IL-10 shRNA. Groups of BALB/c mice were intraperitoneally injected with LPS (25 μg). After two hours, mice received intravenously VLPs-IL-10 shRNA and irrelevant shRNA. The survival rate and survival time of the animals were observed and compared between groups.
Mentions: All mice treated with PBS only or LPS-VLPs-IL-10 shRNA survived in the end of the experiment, but all mice treated with LPS only or LPS-VLPs irrelevant shRNA died after 24 h of treatment. The mice treated with LPS-VLPs-IL-10 shRNA had longer lifespan than those treated with LPS-VLPs irrelevant shRNA or LPS. (Fig. 5)

Bottom Line: In RAW 264.7 cells, IL-10 shRNA was found to reduce IL-10 expression by 85 to 89%, as compared with VLPs alone.IL-10 shRNA did not cross-react with TNF-alpha mRNA or influence the expression of TNF-alpha.In BALB/c mice IL-10 shRNA could reduce 95% of IL-10 secretion.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.

ABSTRACT

Background: RNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Delivering sequences of RNAi in vivo remains a problem. The aim of this study was to use JC virus (JCV) virus-like particles (VLPs) as a vector for delivering RNAi in silencing the cytokine gene of IL-10.

Methods: JCV VLPs were generated by recombinant JCV VP1 protein in yeast expression system. DNA fragment containing IL-10 shRNA was packaged into VLPs by osmotic shock.

Results: In RAW 264.7 cells, IL-10 shRNA was found to reduce IL-10 expression by 85 to 89%, as compared with VLPs alone. IL-10 shRNA did not cross-react with TNF-alpha mRNA or influence the expression of TNF-alpha. In BALB/c mice IL-10 shRNA could reduce 95% of IL-10 secretion. Surprisingly, it also down regulated TNF-alpha expression.

Conclusions: We show for the first time that JCV VLPs empty capsids are competent vectors to deliver RNAi and are nontoxic to cells, suggesting that JCV VLPs is an efficient agent to deliver RNAi in both murine macrophage cells and BALB/c mice. This system provides an efficient means for delivering the RNAi for gene therapy purposes.

Show MeSH
Related in: MedlinePlus