Alba Raul, Baker Andrew H, Nicklin Stuart A
Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, UK.
Methods Mol Biol. 2012;891:55-84. doi: 10.1007/978-1-61779-873-3_4.
Adenoviruses have many attributes, which have made them one of the most widely investigated vectors for gene therapy applications. These include ease of genetic manipulation to produce replication-deficient vectors, ability to readily generate high titer stocks, efficiency of gene delivery into many cell types, and ability to encode large genetic inserts. Recent advances in adenoviral vector engineering have included the ability to genetically manipulate the tropism of the vector by engineering of the major capsid proteins, particularly fiber and hexon. Furthermore, simple replication-deficient adenoviral vectors deleted for expression of a single gene have been complemented by the development of systems in which the majority of adenoviral genes are deleted, generating sophisticated Ad vectors which can mediate sustained transgene expression following a single delivery. This chapter outlines methods for developing simple transgene over expressing Ad vectors and detailed strategies to engineer mutations into the major capsid proteins.
腺病毒具有许多特性，这使其成为基因治疗应用中研究最为广泛的载体之一。这些特性包括易于进行基因操作以产生复制缺陷型载体、能够轻松制备高滴度病毒原液、将基因导入多种细胞类型的效率以及编码大片段遗传插入物的能力。腺病毒载体工程的最新进展包括通过对主要衣壳蛋白（特别是纤维蛋白和六邻体蛋白）进行工程改造来对载体的靶向性进行基因操作的能力。此外，针对单个基因表达进行缺失的简单复制缺陷型腺病毒载体已通过开发大多数腺病毒基因缺失的系统得到补充，从而产生了复杂的腺病毒载体，这些载体在单次递送后能够介导持续的转基因表达。本章概述了开发简单的过表达转基因腺病毒载体的方法以及对主要衣壳蛋白进行突变工程改造的详细策略。
