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The gRNA-miRNA-gRNA Ternary Cassette Combining CRISPR/Cas9 with RNAi Approach Strongly Inhibits Hepatitis B Virus Replication.
- Source :
-
Theranostics [Theranostics] 2017 Jul 22; Vol. 7 (12), pp. 3090-3105. Date of Electronic Publication: 2017 Jul 22 (Print Publication: 2017). - Publication Year :
- 2017
-
Abstract
- The CRISPR/Cas9 system is a novel genome editing technology which has been successfully used to inhibit HBV replication. Here, we described a novel gRNA-microRNA (miRNA)-gRNA ternary cassette driven by a single U6 promoter. With an anti-HBV pri-miR31 mimic integrated between two HBV-specific gRNAs, both gRNAs could be separated from the long transcript of gRNA-miR-HBV-gRNA ternary cassette through Drosha/DGCR8 processing. The results showed that the gRNA-miR-HBV-gRNA ternary cassette could efficiently express two gRNAs and miR-HBV. The optimal length of pri-miRNA flanking sequence in our ternary cassette was determined to be 38 base pairs (bp). Besides, HBV-specific gRNAs and miR-HBV in gRNA-miR-HBV-gRNA ternary cassette could exert a synergistic effect in inhibiting HBV replication and destroying HBV genome in vitro and in vivo . Most importantly, together with RNA interference (RNAi) approach, the HBV-specific gRNAs showed the potent activity on the destruction of HBV covalently closed circular DNA (cccDNA). Since HBV cccDNA is an obstacle for the elimination of chronic HBV infection, the gRNA-miR-HBV-gRNA ternary cassette may be a potential tool for the clearance of HBV cccDNA.<br />Competing Interests: Competing Interests: The authors have declared that no competing interest exists.
- Subjects :
- Antiviral Agents pharmacology
Biological Products pharmacology
Cell Line
DNA, Circular metabolism
DNA, Viral metabolism
Hepatitis B virus genetics
Hepatocytes virology
Humans
MicroRNAs metabolism
RNA, Guide, CRISPR-Cas Systems metabolism
CRISPR-Cas Systems
Gene Knockdown Techniques methods
Gene Targeting methods
Hepatitis B virus physiology
RNA Interference
Virus Replication
Subjects
Details
- Language :
- English
- ISSN :
- 1838-7640
- Volume :
- 7
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Theranostics
- Publication Type :
- Academic Journal
- Accession number :
- 28839466
- Full Text :
- https://doi.org/10.7150/thno.18114