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标题: 通过DNA-PKcs抑制剂NU7026抑制NHEJ途径防止由CRISPR / Cas9切割的HBV [打印本页]

作者: StephenW    时间: 2019-2-17 13:29     标题: 通过DNA-PKcs抑制剂NU7026抑制NHEJ途径防止由CRISPR / Cas9切割的HBV

Sci Rep. 2019 Feb 12;9(1):1847. doi: 10.1038/s41598-019-38526-6.
Suppressing the NHEJ pathway by DNA-PKcs inhibitor NU7026 prevents degradation of HBV cccDNA cleaved by CRISPR/Cas9.
Kostyushev D1, Kostyusheva A2, Brezgin S2,3, Zarifyan D2, Utkina A2, Goptar I2,4, Chulanov V2,5.
Author information

1
    Central Research Institute of Epidemiology, Viral Hepatitis, Moscow, 111123, Russian Federation. [email protected].
2
    Central Research Institute of Epidemiology, Viral Hepatitis, Moscow, 111123, Russian Federation.
3
    Institute of Immunology, Federal Medical Biological Agency, Moscow, 115478, Russian Federation.
4
    Izmerov Research Institute of Occupational Health, Gene Engineering and Biotechnology, Moscow, 105275, Russian Federation.
5
    I.M. Sechenov First Moscow State Medical University, Infectious Diseases, Moscow, 119146, Russian Federation.

Abstract

Chronic hepatitis B is a severe liver disease caused by hepatitis B virus (HBV) infection. Covalently closed circular DNA (cccDNA), a super-spiralized, double-stranded form of the HBV genome, is the major determinant of viral persistence. CRISPR/Cas9 nucleases have been recently shown to introduce double-stranded DNA breaks into HBV cccDNA. The inflicted damage results predominantly in erroneous repair of cccDNA by non-homologous end-joining (NHEJ). NHEJ has been suggested to enhance anti-HBV activity of CRISPR/Cas9 and increase cccDNA mutation. In this study, we assessed anti-HBV activity of CRISPR/Cas9 and cccDNA repair outcomes in an altered NHEJ/HR environment. NU7026, a strong inhibitor of NHEJ, prevented CRISPR/Cas9-mediated degradation of cccDNA and resulted in frequent on-target deletions. We conclude that CRISPR/Cas9 is a highly effective tool to degrade cccDNA and first demonstrate that inhibiting NHEJ impairs cccDNA degradation.

PMID:
    30755668
DOI:
    10.1038/s41598-019-38526-6
作者: StephenW    时间: 2019-2-17 13:29

SciRep.2019年2月12日; 9(1):1847。 doi:10.1038 / s41598-019-38526-6。
通过DNA-PKcs抑制剂NU7026抑制NHEJ途径防止由CRISPR / Cas9切割的HBV cccDNA的降解。
Kostyushev D1,Kostyusheva A2,Brezgin S2,3,Zarifyan D2,Utkina A2,Goptar I2,4,Chulanov V2,5。
作者信息

1
    中央流行病学研究所,病毒性肝炎,莫斯科,111123,俄罗斯联邦。 [email protected]
2
    中央流行病学研究所,病毒性肝炎,莫斯科,111123,俄罗斯联邦。
3
    联邦医学生物机构免疫学研究所,莫斯科,115478,俄罗斯联邦。
4
    Izmerov职业健康研究所,基因工程和生物技术,莫斯科,105275,俄罗斯联邦。

    I.M. Sechenov第一莫斯科国立医科大学,传染病,莫斯科,119146,俄罗斯联邦。

抽象

慢性乙型肝炎是由乙型肝炎病毒(HBV)感染引起的严重肝病。共价闭合环状DNA(cccDNA)是HBV基因组的超螺旋双链形式,是病毒持续性的主要决定因素。最近已显示CRISPR / Cas9核酸酶将双链DNA断裂引入HBV cccDNA。造成的损害主要导致通过非同源末端连接(NHEJ)错误修复cccDNA。已建议NHEJ增强CRISPR / Cas9的抗HBV活性并增加cccDNA突变。在这项研究中,我们评估了改变的NHEJ / HR环境中CRISPR / Cas9的抗HBV活性和cccDNA修复结果。 NU7026是NHEJ的强抑制剂,可阻止CRISPR / Cas9介导的cccDNA降解,并导致频繁的靶向缺失。我们得出结论,CRISPR / Cas9是降解cccDNA的高效工具,并首次证明抑制NHEJ会损害cccDNA降解。

结论:
    30755668
DOI:
    10.1038 / s41598-019-38526-6
作者: StephenW    时间: 2019-2-17 13:30

https://www.nature.com/articles/s41598-019-38526-6
作者: 齐欢畅    时间: 2019-2-17 19:31

我们得出结论,CRISPR / Cas9是降解cccDNA的高效工具,并首次证明抑制NHEJ会损害cccDNA降解。

作者: fuke    时间: 2019-2-17 22:09

回复 齐欢畅 的帖子

进入临床就知道效果了,这个是治本的药。
作者: pourvivre    时间: 2019-2-18 11:45

跟进




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