文献类型：期刊文献

英文题名：Inactivation of the key ORFs of HBV for antiviral therapy by non-cleavage base editing

作者：Qin, Bo[1,2];Shen, Shu[1,2];Chen, Hao[1,2];Wang, Yiying[1,2];Ding, Jinlong[1,2];Ding, Jiefeng[1,2]

机构：[1]Shaoxing Matern & Child Hlth Care Hosp, Shaoxing, Peoples R China;[2]Shaoxing Univ, Obstet & Gynecol Hosp, Shaoxing, Peoples R China

年份：2025

卷号：205

外文期刊名：MICROBIAL PATHOGENESIS

收录：SCI-EXPANDED(收录号：WOS:001499639000002)、、Scopus(收录号：2-s2.0-105005512909)、WOS

基金：This work was supported by the Science Technology Department of Zhejiang Province, China (LGF22H190009, TGY24H190011) , the Health Commission of Zhejiang Province, China (2022KY411) , the Traditional Chinese Medicine Project of Zhejiang Province, China (2024ZL1140) , and open-ended foundation of Shaoxing health commission, Zhejiang Province of China (2023SSY002) , the Wu Jieping Medical Foundation Specialized Research Grant on Blood Biomarkers Tracing for Assessment and Decision-Making in Critical and Emergency Conditions (320.6750.2024-23-05) .

语种：英文

外文关键词：Hepatitis B virus (HBV); Clustered regularly interspaced short; palindromic repeats (CRISPR); Double-strand breaks (DSBs); Adenine base editors (ABEs); Cytosine base editors (CBEs)

外文摘要：Objectives: Hepatitis B virus (HBV) infection is the key cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Currently available anti-HBV drugs are more or less defective owing to the unremovable covalently closed circular DNA (cccDNA). Thus, CRISPR/Cas9 is a promising therapeutic strategy for anti-HBV therapy. Double-strand breaks (DSBs) and uncontrolled genomic rearrangements occur inevitably. In this study, we aimed to use base editors to control HBV infection. Methods: Base editors precisely instal targeted point mutations without requiring DSBs or donor DNA templates, and without relying on homology-directed repair (HDR) or nonhomologous end joining (NHEJ). Adenine base editors (ABEs) and cytosine base editors (CBEs) catalyse A center dot T to G center dot C and C center dot G to T center dot A conversions, respectively. In this study, to control HBV replication by modifying and inactivating key HBV genes, recently developed CRISPR/Cas-mediated SpRY-ABE8e and CBE4-max were utilised to falsify and invalidate the ATG initiation codons of the S, Pre-S1, PreS2, C, Pre-C, X, and P genes. Results: The ATG initiation codons of HBV genes were edited by ABE/CBE. The expected point mutations were successfully introduced, resulting in the simultaneous suppression of HBV antigen expression and replication to varying degrees. Conclusions: Our study focused on clearing HBV using base and provided experimental and theoretical evidence for the treatment of chronic HBV infection. Thus, base editing is a potential strategy for curing CHB by permanently inactivating the integrated DNA and cccDNA without using DSBs.