| Title: |
CRISPR-Cas13b-mediated suppression of HBV replication and protein expression |
| Authors: |
McCoullough, LC; Fareh, M; Hu, W; Sozzi, V; Makhlouf, C; Droungas, Y; Lee, CL; Takawy, M; Fabb, SA; Payne, TJ; Pouton, CW; Netter, HJ; Lewin, SR; Purcell, DFJ; Holmes, JA; Trapani, JA; Littlejohn, M; Revill, PA |
| Publisher Information: |
Elsevier |
| Publication Year: |
2024 |
| Collection: |
The University of Melbourne: Digital Repository |
| Description: |
BACKGROUND & AIMS: New antiviral approaches that target multiple aspects of the HBV replication cycle to improve rates of functional cure are urgently required. HBV RNA represents a novel therapeutic target. Here, we programmed CRISPR-Cas13b endonuclease to specifically target the HBV pregenomic RNA and viral mRNAs in a novel approach to reduce HBV replication and protein expression. METHODS: Cas13b CRISPR RNAs (crRNAs) were designed to target multiple regions of HBV pregenomic RNA. Mammalian cells transfected with replication competent wild-type HBV DNA of different genotypes, a HBV-expressing stable cell line, a HBV infection model and a hepatitis B surface antigen (HBsAg)-expressing stable cell line were transfected with PspCas13b-BFP (blue fluorescent protein) and crRNA plasmids, and the impact on HBV replication and protein expression was measured. Wild-type HBV DNA, PspCas13b-BFP and crRNA plasmids were simultaneously hydrodynamically injected into mice, and serum HBsAg was measured. PspCas13b mRNA and crRNA were also delivered to a HBsAg-expressing stable cell line via lipid nanoparticles and the impact on secreted HBsAg determined. RESULTS: Our HBV-targeting crRNAs strongly suppressed HBV replication and protein expression in mammalian cells by up to 96% (p |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| ISSN: |
0168-8278 |
| Relation: |
NHMRC/1145977; https://hdl.handle.net/11343/349191 |
| Availability: |
https://hdl.handle.net/11343/349191 |
| Rights: |
https://creativecommons.org/licenses/by/4.0 ; CC BY |
| Accession Number: |
edsbas.8AB3DC4E |
| Database: |
BASE |