用于设计者核酸酶表达的体外转录MRNA：作为治疗慢性HBV感染

StephenW

In Vitro Transcribed MRNA for Expression of Designer Nucleases: Advantages as a Novel Therapeutic for the Management of Chronic HBV Infection
Abdullah Ely  1 , Prashika Singh  1 , Tiffany S Smith  1 , Patrick Arbuthnot  2
Affiliations
Affiliations

    1
    Wits/SAMRC Antiviral Gene Therapy Research Unit, Department of Molecular Medicine and Haematology, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, South Africa.
    2
    Wits/SAMRC Antiviral Gene Therapy Research Unit, Department of Molecular Medicine and Haematology, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, South Africa. Electronic address: [email protected].

    PMID: 32485207 DOI: 10.1016/j.addr.2020.05.010

Abstract

Chronic infection with the hepatitis B virus (HBV) remains a significant worldwide medical problem. While diseases caused by HIV infection, tuberculosis and malaria are on the decline, new cases of chronic hepatitis B are on the rise. Because often fatal complications of cirrhosis and hepatocellular carcinoma are associated with chronic hepatitis B, the need for a cure is as urgent as ever. Currently licensed therapeutics fail to eradicate the virus and this is attributable to persistence of the viral replication intermediate comprising covalently closed circular DNA (cccDNA). Elimination or inactivation of the viral cccDNA is thus a goal of research aimed at hepatitis B cure. Ability to engineer nucleases that are capable of specific cleavage of a DNA sequence now provides the means to disable cccDNA permanently. The scientific literature is replete with many examples of using designer zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and RNA-guided endonucleases (RGENs) to inactivate HBV. However, important concerns about safety, dose control and efficient delivery need to be addressed before the technology is employed in a clinical setting. Use of in vitro transcribed mRNA to express therapeutic gene editors goes some way to overcoming these concerns. The labile nature of RNA limits off-target effects and enables dose control. Compatibility with hepatotropic non-viral vectors is convenient for the large scale preparation that will be required for advancing gene editing as a mode of curing chronic hepatitis B.

Keywords: CRISPR/Cas; Non-viral vectors; TALENs; ZFNs; cccDNA.

Copyright © 2020. Published by Elsevier B.V.

StephenW

用于设计者核酸酶表达的体外转录MRNA：作为治疗慢性HBV感染的新型疗法的优势
Abdullah Ely 1，Prashika Singh 1，Tiffany S Smith 1，Patrick Arbuthnot 2
隶属关系
隶属关系

1个
南非威特沃兹兰德大学卫生学院病理学学院分子医学和血液学系Wits / SAMRC抗病毒基因治疗研究室。
2
南非威特沃兹兰德大学卫生学院病理学学院分子医学与血液学系Wits / SAMRC抗病毒基因治疗研究室。电子地址：[email protected]。

PMID：32485207 DOI：10.1016 / j.addr.2020.05.010

抽象

乙型肝炎病毒（HBV）的慢性感染仍然是世界范围内的重大医学问题。由艾滋病毒感染，结核病和疟疾引起的疾病正在减少，而慢性乙型肝炎的新病例正在增加。由于肝硬化和肝细胞癌的致命并发症通常与慢性乙型肝炎有关，因此对治疗的需求与以往一样迫切。当前获得许可的治疗剂不能根除该病毒，这归因于包含共价封闭的环状DNA（cccDNA）的病毒复制中间体的持久性。因此，消除或灭活病毒cccDNA是针对乙型肝炎的研究目标。现在，能够特异性切割DNA序列的核酸酶的工程设计能力提供了永久禁用cccDNA的方法。科学文献中充斥着许多使用设计师锌指核酸酶（ZFN），转录激活因子样效应核酸酶（TALEN）和RNA引导d核酸内切酶（RGEN）灭活HBV的例子。但是，在将该技术用于临床之前，必须解决与安全性，剂量控制和有效递送有关的重要问题。使用体外转录的mRNA表达治疗性基因编辑器可以克服这些问题。 RNA的不稳定性质限制了脱靶效应并可以控制剂量。与促肝非病毒载体的相容性便于大规模制备，这对于推进基因编辑作为治愈慢性乙型肝炎的一种方式将是必需的。

关键词：CRISPR / Cas；非病毒载体； TALEN； ZFN； cccDNA。

版权所有©2020。由Elsevier B.V.发布。

lancas

感谢楼主分享啊~这么好的板块就给这些人毁了