CRISPR/Cas9 delivery by NIR-responsive biomimetic nanoparticles for targeted HBV therapy

Dan Wang,Hua Tang,Qing Yang,Ling Chen,Ailong Huang,Chengbi Li,Quanxin Long

doi:10.1186/s12951-021-01233-4

Dan Wang, Hua Tang + Show 5 more

Open Access

https://doi.org/10.1186/s12951-021-01233-4

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Abstract

BackgroundCurrently, there are no curative drugs for hepatitis B virus (HBV). Complete elimination of HBV covalently closed circular DNA (cccDNA) is key to the complete cure of hepatitis B virus infection. The CRISPR/Cas9 system can directly destroy HBV cccDNA. However, a CRISPR/Cas9 delivery system with low immunogenicity and high efficiency has not yet been established. Moreover, effective implementation of precise remote spatiotemporal operations in CRISPR/Cas9 is a major limitation.ResultsIn this work, we designed NIR-responsive biomimetic nanoparticles (UCNPs-Cas9@CM), which could effectively deliver Cas9 RNP to achieve effective genome editing for HBV therapy. HBsAg, HBeAg, HBV pgRNA and HBV DNA along with cccDNA in HBV-infected cells were found to be inhibited. These findings were confirmed in HBV-Tg mice, which did not exhibit significant cytotoxicity and minimal off-target DNA damage.ConclusionsThe UCNPs-based biomimetic nanoplatforms achieved the inhibition of HBV replication via CRISPR therapy and it is a potential system for efficient treatment of human HBV diseases.Graphical

Highlights

There are no curative drugs for hepatitis B virus (HBV)
Transmission electron microscopy (TEM) mapping confirmed the composite structure of upconversion nanoparticles (UCNPs) (Fig. 1D)
Due to the strong affinity between avidin and biotin, PC Biotin-NHS Ester (PCB) could bind UCNPs-Avidin to form UCNPs-Avidin/ PCB nanoparticles, which were covalently conjugated with Cas9 proteins and incubated with Single guide RNA (sgRNA) to generate

Summary

Introduction

There are no curative drugs for hepatitis B virus (HBV). Complete elimination of HBV covalently closed circular DNA (cccDNA) is key to the complete cure of hepatitis B virus infection. Lengthy treatment is associated with drug resistance mutations and a risk of Hepatic Fibrosis/Cirrhosis These drugs cannot target or eliminate HBV covalently closed circular DNA (cccDNA), which inhibits complete neutralization of chronic hepatitis B (CHB) infections [2, 3]. Elimination of cccDNA through epigenetic modifications, such as histone modifications and methylation of HBV cccDNA, or gene-editing therapies, such as Clustered Regularly Inter Spaced Palindromic Repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR/ Cas9) [4,5,6]. Among these strategies, CRISPR/Cas is currently the most promising therapeutic means [7,8,9]

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