[CRISPR/Cas9 核糖核蛋白複合物對體內乙型肝炎病毒模型的抗病

StephenW

[CRISPR/Cas9 核糖核蛋白複合物對體內乙型肝炎病毒模型的抗病毒活性]
[俄語文章]
A P Kostyusheva 1 , S A Brezgin 1 2 , N I Ponomareva 1 2 , I A Goptar 3 , A V Nikiforova 3 , V I Gegechkori 1 , V B Poluektova 1 , K A Turkadze 1 , A E Sudina 4 , V P Chulanov 1 2 4 , D S 2 Kostyushev 1
隸屬關係

     PMID：36475475 DOI：10.31857/S002689842206012X

抽象的

慢性乙型肝炎（CHB）是由乙型肝炎病毒（HBV）感染引起的。 這種疾病是全球健康的關鍵問題。 現代治療方法不能從受感染的細胞中完全清除 HBV，也不能治愈慢性感染。 CRISPR/Cas9 位點特異性核酸酶系統可以有效切割不針對包括病毒基因組在內的 DNA。 HBV 基因組主要形式的切割，即共價閉合環狀 DNA (cccDNA)，導致病毒複製和 cccDNA 降解或突變失活的強烈減少。 基於 CRISPR/Cas9 的方法是實現 CHB“殺菌”治療（即從體內完全清除病毒）的最有希望的方法之一。 在這裡，體內的 HBV 小鼠模型已用於分析針對 HBV 基因組的高特異性 Cas9 蛋白和 sgRNA 的抗病毒活性。 我們發現，早在實驗開始後 48 小時，單次注射 CRISPR/Cas9 的短壽命核糖核蛋白複合物就會導緻小鼠血清和肝臟中的 HBV DNA 水平降低約 10 倍。 已發現剩餘的 HBV DNA 具有罕見的插入缺失突變。 基於 CRISPR/Cas9 核糖核蛋白複合物開發治療 CHB 的新型抗病毒藥物可以顯著縮短 CHB 治療的持續時間，並有可能實現完全消除病毒感染。

關鍵詞：NGS； 抗病毒藥物； 刪除； 基因編輯； 基因組技術； 插入。

StephenW

[Antiviral Activity of CRISPR/Cas9 Ribonucleoprotein Complexes on a Hepatitis B Virus Model In Vivo]
[Article in Russian]
A P Kostyusheva  1 , S A Brezgin  1   2 , N I Ponomareva  1   2 , I A Goptar  3 , A V Nikiforova  3 , V I Gegechkori  1 , V B Poluektova  1 , K A Turkadze  1 , A E Sudina  4 , V P Chulanov  1   2   4 , D S Kostyushev  1   2   5
Affiliations

    PMID: 36475475 DOI: 10.31857/S002689842206012X

Abstract

Chronic hepatitis B (CHB) is caused by hepatitis B virus (HBV) infection. This disease is a key issue for global health. Modern methods of therapy do not completely eliminate HBV from infected cells and do not cure chronic infection. The CRISPR/Cas9 systems of site-specific nucleases can effectively cleave do not target DNA including viral genomes. The cleavage of the major form of the HBV genome, i.e., covalently closed circular DNA (cccDNA), leads to a robust reduction in viral replication and degradation or mutational inactivation of cccDNA. CRISPR/Cas9-based approaches are one of the most promising ways to achieve a 'sterilizing' cure of CHB, i.e., complete elimination of the virus from the body. Here, the HBV mouse model in vivo has been used to analyze the antiviral activity of the high-specific Cas9 protein and sgRNA targeting HBV genome. We have found that a single injection of short-lived ribonucleoprotein complexes of CRISPR/Cas9 results in a ~10-fold reduction in HBV DNA levels in the serum and liver of mice as early as 48 h after the start of the experiment. The remaining HBV DNAs have been found to harbor rare indel mutations. Developing new antivirals for treating CHB based on CRISPR/Cas9 ribonucleoprotein complexes could substantially reduce the duration of CHB therapy and, potentially, achieve complete elimination of viral infection.

Keywords: NGS; antiviral drugs; deletions; gene editing; genome technologies; insertions.