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RNA Interference inhibits hepatitis B virus of different genotypes in vitro and in vivo.

Zhang YL, Cheng T, Cai YJ, Yuan Q, Liu C, Zhang T, Xia DZ, Li RY, Yang LW, Wang YB, Yeo AE, Shih JW, Zhang J, Xia NS - BMC Microbiol. (2010)

Bottom Line: Small interfering RNA (siRNA) can be a potential new tool for HBV therapy.Given the high heterogeneity of HBV strains and the sensitivity towards sequences changes of siRNA, finding a potent siRNA inhibitor against the conservative site on the HBV genome is essential to ensure a therapeutic application.No unusual cytotoxicity or off-target effects were noted.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science, Xiamen University, Xiamen, Fujian Province, China.

ABSTRACT

Background: Hepatitis B virus (HBV) infection increases the risk of liver disease and hepatocellular carcinoma. Small interfering RNA (siRNA) can be a potential new tool for HBV therapy. Given the high heterogeneity of HBV strains and the sensitivity towards sequences changes of siRNA, finding a potent siRNA inhibitor against the conservative site on the HBV genome is essential to ensure a therapeutic application.

Results: Forty short hairpin RNA (shRNA) expression plasmids were constructed to target conserved regions among nine HBV genotypes. HBV 1.3-fold genome plasmids carrying various genotypes were co-transfected with shRNA plasmids into either Huh7 cells or mice. The levels of various viral markers were examined to assess the anti-HBV efficacy of siRNA. Four (B245, B376, B1581 and B1789) were found with the ability to potently inhibit HBV RNA, DNA, surface antigen (HBsAg), e antigen (HBeAg) and core antigen (HBcAg) expression in HBV genotypes A, B, C, D and I (a newly identified genotype) in Huh7 cells and in mice. No unusual cytotoxicity or off-target effects were noted.

Conclusions: Such siRNA suggests an alternate way of inhibiting various HBV genotypes in vitro and in vivo, promising advances in the treatment of HBV.

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Related in: MedlinePlus

SiRNAs inhibit RNA and DNA expression of HBV strains with different genotypes in Huh7 cells. The histogram show the cytoplasmic HBV pg/pc RNA levels (A, B, C, D, E) and extracellular HBV DNA (F, G, H, I, J) of five HBV strains with genotypes Ae(N10), Ba(C4371), C1(Y1021), D1(Y10) and I1(W29) in treated shRNA plasmids, treated pSUPER vector, and non-treated Huh7 cells.
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Figure 3: SiRNAs inhibit RNA and DNA expression of HBV strains with different genotypes in Huh7 cells. The histogram show the cytoplasmic HBV pg/pc RNA levels (A, B, C, D, E) and extracellular HBV DNA (F, G, H, I, J) of five HBV strains with genotypes Ae(N10), Ba(C4371), C1(Y1021), D1(Y10) and I1(W29) in treated shRNA plasmids, treated pSUPER vector, and non-treated Huh7 cells.

Mentions: The levels of cytoplasmic HBV pg/pc RNA (3.5 kb) and HBV DNA in cultured supernatants were determined by realtime RT-PCR/PCR and presented in Figure 3. The pg/pc RNA level of five HBV strains with different genotypes were reduced by 58%~93% in B245(69%~93%), B376(59%~91%), B1581(67%~90%) and B1789(58%~88%) treatments, while the HBV DNA level observed in supernatants was decreased by 77%~99% in these shRNA plasmid treatments (B245: 83%~99%, B376: 79%~99%, B1581:88%~98%, B1789: 77%~99%).


RNA Interference inhibits hepatitis B virus of different genotypes in vitro and in vivo.

Zhang YL, Cheng T, Cai YJ, Yuan Q, Liu C, Zhang T, Xia DZ, Li RY, Yang LW, Wang YB, Yeo AE, Shih JW, Zhang J, Xia NS - BMC Microbiol. (2010)

SiRNAs inhibit RNA and DNA expression of HBV strains with different genotypes in Huh7 cells. The histogram show the cytoplasmic HBV pg/pc RNA levels (A, B, C, D, E) and extracellular HBV DNA (F, G, H, I, J) of five HBV strains with genotypes Ae(N10), Ba(C4371), C1(Y1021), D1(Y10) and I1(W29) in treated shRNA plasmids, treated pSUPER vector, and non-treated Huh7 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927532&req=5

Figure 3: SiRNAs inhibit RNA and DNA expression of HBV strains with different genotypes in Huh7 cells. The histogram show the cytoplasmic HBV pg/pc RNA levels (A, B, C, D, E) and extracellular HBV DNA (F, G, H, I, J) of five HBV strains with genotypes Ae(N10), Ba(C4371), C1(Y1021), D1(Y10) and I1(W29) in treated shRNA plasmids, treated pSUPER vector, and non-treated Huh7 cells.
Mentions: The levels of cytoplasmic HBV pg/pc RNA (3.5 kb) and HBV DNA in cultured supernatants were determined by realtime RT-PCR/PCR and presented in Figure 3. The pg/pc RNA level of five HBV strains with different genotypes were reduced by 58%~93% in B245(69%~93%), B376(59%~91%), B1581(67%~90%) and B1789(58%~88%) treatments, while the HBV DNA level observed in supernatants was decreased by 77%~99% in these shRNA plasmid treatments (B245: 83%~99%, B376: 79%~99%, B1581:88%~98%, B1789: 77%~99%).

Bottom Line: Small interfering RNA (siRNA) can be a potential new tool for HBV therapy.Given the high heterogeneity of HBV strains and the sensitivity towards sequences changes of siRNA, finding a potent siRNA inhibitor against the conservative site on the HBV genome is essential to ensure a therapeutic application.No unusual cytotoxicity or off-target effects were noted.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science, Xiamen University, Xiamen, Fujian Province, China.

ABSTRACT

Background: Hepatitis B virus (HBV) infection increases the risk of liver disease and hepatocellular carcinoma. Small interfering RNA (siRNA) can be a potential new tool for HBV therapy. Given the high heterogeneity of HBV strains and the sensitivity towards sequences changes of siRNA, finding a potent siRNA inhibitor against the conservative site on the HBV genome is essential to ensure a therapeutic application.

Results: Forty short hairpin RNA (shRNA) expression plasmids were constructed to target conserved regions among nine HBV genotypes. HBV 1.3-fold genome plasmids carrying various genotypes were co-transfected with shRNA plasmids into either Huh7 cells or mice. The levels of various viral markers were examined to assess the anti-HBV efficacy of siRNA. Four (B245, B376, B1581 and B1789) were found with the ability to potently inhibit HBV RNA, DNA, surface antigen (HBsAg), e antigen (HBeAg) and core antigen (HBcAg) expression in HBV genotypes A, B, C, D and I (a newly identified genotype) in Huh7 cells and in mice. No unusual cytotoxicity or off-target effects were noted.

Conclusions: Such siRNA suggests an alternate way of inhibiting various HBV genotypes in vitro and in vivo, promising advances in the treatment of HBV.

Show MeSH
Related in: MedlinePlus