Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells.
External guide sequences (EGSs) are RNA molecules that consist of a sequence complementary to a target mRNA and recruit intracellular ribonuclease P (RNase P), a tRNA processing enzyme, for specific degradation of the target mRNA. We have previously used an in vitro selection procedure to generate E...
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3680410?pdf=render |
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| author | Zhigang Zhang Gia-Phong Vu Hao Gong Chuan Xia Yuan-Chuan Chen Fenyong Liu Jianguo Wu Sangwei Lu |
| author_facet | Zhigang Zhang Gia-Phong Vu Hao Gong Chuan Xia Yuan-Chuan Chen Fenyong Liu Jianguo Wu Sangwei Lu |
| author_sort | Zhigang Zhang |
| collection | DOAJ |
| description | External guide sequences (EGSs) are RNA molecules that consist of a sequence complementary to a target mRNA and recruit intracellular ribonuclease P (RNase P), a tRNA processing enzyme, for specific degradation of the target mRNA. We have previously used an in vitro selection procedure to generate EGS variants that efficiently induce human RNase P to cleave a target mRNA in vitro. In this study, we constructed EGSs from a variant to target the overlapping region of the S mRNA, pre-S/L mRNA, and pregenomic RNA (pgRNA) of hepatitis B virus (HBV), which are essential for viral replication and infection. The EGS variant was about 50-fold more efficient in inducing human RNase P to cleave the mRNA in vitro than the EGS derived from a natural tRNA. Following Salmonella-mediated gene delivery, the EGSs were expressed in cultured HBV-carrying cells. A reduction of about 97% and 75% in the level of HBV RNAs and proteins and an inhibition of about 6,000- and 130-fold in the levels of capsid-associated HBV DNA were observed in cells treated with Salmonella vectors carrying the expression cassette for the variant and the tRNA-derived EGS, respectively. Our study provides direct evidence that the EGS variant is more effective in blocking HBV gene expression and DNA replication than the tRNA-derived EGS. Furthermore, these results demonstrate the feasibility of developing Salmonella-mediated gene delivery of highly active EGS RNA variants as a novel approach for gene-targeting applications such as anti-HBV therapy. |
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| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T01:06:34Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-4c43dd17f4e44f838c5de649913a64a92022-12-22T01:26:09ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0186e6526810.1371/journal.pone.0065268Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells.Zhigang ZhangGia-Phong VuHao GongChuan XiaYuan-Chuan ChenFenyong LiuJianguo WuSangwei LuExternal guide sequences (EGSs) are RNA molecules that consist of a sequence complementary to a target mRNA and recruit intracellular ribonuclease P (RNase P), a tRNA processing enzyme, for specific degradation of the target mRNA. We have previously used an in vitro selection procedure to generate EGS variants that efficiently induce human RNase P to cleave a target mRNA in vitro. In this study, we constructed EGSs from a variant to target the overlapping region of the S mRNA, pre-S/L mRNA, and pregenomic RNA (pgRNA) of hepatitis B virus (HBV), which are essential for viral replication and infection. The EGS variant was about 50-fold more efficient in inducing human RNase P to cleave the mRNA in vitro than the EGS derived from a natural tRNA. Following Salmonella-mediated gene delivery, the EGSs were expressed in cultured HBV-carrying cells. A reduction of about 97% and 75% in the level of HBV RNAs and proteins and an inhibition of about 6,000- and 130-fold in the levels of capsid-associated HBV DNA were observed in cells treated with Salmonella vectors carrying the expression cassette for the variant and the tRNA-derived EGS, respectively. Our study provides direct evidence that the EGS variant is more effective in blocking HBV gene expression and DNA replication than the tRNA-derived EGS. Furthermore, these results demonstrate the feasibility of developing Salmonella-mediated gene delivery of highly active EGS RNA variants as a novel approach for gene-targeting applications such as anti-HBV therapy.http://europepmc.org/articles/PMC3680410?pdf=render |
| spellingShingle | Zhigang Zhang Gia-Phong Vu Hao Gong Chuan Xia Yuan-Chuan Chen Fenyong Liu Jianguo Wu Sangwei Lu Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. PLoS ONE |
| title | Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. |
| title_full | Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. |
| title_fullStr | Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. |
| title_full_unstemmed | Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. |
| title_short | Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. |
| title_sort | engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis b virus in cultured cells |
| url | http://europepmc.org/articles/PMC3680410?pdf=render |
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