利用Illumina新一代测序技术对腺相关病毒质粒进行准确、快速的序列分析。

Q1 Immunology and Microbiology Human Gene Therapy Methods Pub Date : 2018-10-01 Epub Date: 2018-08-22 DOI:10.1089/hgtb.2018.037

Alexei Saveliev, Juan Liu, Mingyao Li, Lee Hirata, Caitlin Latshaw, Jia Zhang, James M Wilson

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引用次数: 5

摘要

质粒DNA的序列验证是一个至关重要的质量控制步骤，必须在通过质粒转染包装腺相关病毒(AAV)载体之前进行。AAV顺式质粒对序列分析提出了独特的挑战，因为它们含有反向末端重复序列，并且易于序列重排。建立了一种准确、快速的新一代测序方法，可以在3.5天内分析AAV顺式质粒的全长序列。本文描述了一种能够可靠地检测和鉴定AAV顺式质粒中常见的序列变异的位置和频率的分步方案。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Accurate and Rapid Sequence Analysis of Adeno-Associated Virus Plasmids by Illumina Next-Generation Sequencing.

Sequence validation of plasmid DNA is a crucial quality control step that must occur prior to adeno-associated virus (AAV) vector packaging through plasmid transfection. AAV cis-plasmids present unique challenges to sequence analysis, as they contain inverted terminal repeats and are prone to sequence rearrangements. An accurate and rapid next-generation sequencing approach has been established to analyze full-length sequences of AAV cis-plasmids within 3.5 days. Here, a step-by-step protocol is described that can reliably detect and identify the location and frequency of sequence variants commonly observed in AAV cis-plasmids.

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来源期刊

Human Gene Therapy Methods BIOTECHNOLOGY & APPLIED MICROBIOLOGY-GENETICS & HEREDITY

CiteScore

5.80

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases. The Journal is divided into three parts. Human Gene Therapy, the flagship, is published 12 times per year. HGT Methods, a bimonthly journal, focuses on the applications of gene therapy to product testing and development. HGT Clinical Development, a quarterly journal, serves as a venue for publishing data relevant to the regulatory review and commercial development of cell and gene therapy products.