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Review
. 2024 Oct 2;16(10):1565.
doi: 10.3390/v16101565.

Applications of CRISPR/Cas as a Toolbox for Hepatitis B Virus Detection and Therapeutics

Anuj Kumar  1   2 , Emmanuel Combe  1   2 , Léa Mougené  1   2 , Fabien Zoulim  1   2   3   4 , Barbara Testoni  1   2
Affiliations
Review

Applications of CRISPR/Cas as a Toolbox for Hepatitis B Virus Detection and Therapeutics

Anuj Kumar et al. Viruses. .

Abstract

Hepatitis B virus (HBV) infection remains a significant global health challenge, leading to chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Covalently closed circular DNA (cccDNA) and integrated HBV DNA are pivotal in maintaining viral persistence. Recent advances in CRISPR/Cas technology offer innovative strategies to inhibit HBV by directly targeting both cccDNA and integrated HBV DNA or indirectly by degrading HBV RNAs or targeting host proteins. This review provides a comprehensive overview of the latest advancements in using CRISPR/Cas to inhibit HBV, with a special highlight on newer non-double-strand (non-DSB) break approaches. Beyond the canonical use of CRISPR/Cas for target inhibition, we discuss additional applications, including HBV diagnosis and developing models to understand cccDNA biology, highlighting the diverse use of this technology in the HBV field.

Keywords: CRISPR/Cas; HBV; cccDNA; diagnostics; gene editing.

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Conflict of interest statement

A.K., E.C., and L.M. declare no conflicts of interest. F.Z. and B.T. received grants from Assembly Biosciences, Bluejay Therapeutics, Beam Therapeutics, and JnJ. F.Z. conducted consulting activities with Assembly Biosciences, Bluejay Therapeutics, and GSK.

Figures

Figure 1
Figure 1
Schematic representation of the HBV transcripts and ORFs expressed from (A) cccDNA and from (B) the integrated HBV DNA. These HBV genomes are crucial for HBV chronicity, and they serve as targets for CRISPR/Cas9 approaches (pro: promoter; PreC: precore; DR: direct repeat; PAS: polyadenylation signal; Enh: enhancer) (adapted from [5,11,12,14]). Image created with BioRender.com.
Figure 2
Figure 2
CRISPR/Cas-based approaches offer potential for both detecting and targeting cccDNA and integrated HBV DNA, which play crucial role in HBV chronicity. While Cas9 nuclease, base editors, and epigenetic editors can directly target viral genomes, Cas13b functions by targeting HBV RNAs. For detection purposes, Cas12 and Cas13 can be employed to detect HBV DNA or RNA, respectively. Although this figure primarily focuses on cccDNA and integrated DNA, Cas12 can also detect other HBV DNA species, including rcDNA. Image created with BioRender.com.
Figure 3
Figure 3
Detection of HBV using Cas12 and Cas13 can be achieved through various readout methods, including fluorescence detection, a lateral-flow immunochromatographic paper-strip assay, electrochemiluminescence, colorimetry, surface-enhanced Raman spectroscopy (SERS), and a personal glucose meter (PGM) (adapted from [98,112,115]). Image created with BioRender.com.
See this image and copyright information in PMC

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