DNA改组在腺相关病毒定向进化中的发展及其应用  

Development and application of DNA shuffling in the directed evolution of adeno-associated virus

作　　者：潘素晶[1] 徐增辉[2] 张悦[1] 钱其军[1,2] 

机构地区：[1]浙江理工大学生命科学学院、新元医学与生物技术研究所,杭州310018 [2]中国人民解放军第二军医大学、东方肝胆外科医院基因病毒治疗实验室,上海200438

出　　处：《科学通报》2013年第5期411-418,共8页Chinese Science Bulletin

基　　金：国家自然科学基金(81071850)资助

摘　　要：DNA改组(DNA shuffling)是一种高通量的突变、筛选技术,自1994年提出以来,经过了几十年的发展,其在DNA、酶和药物蛋白等的改造上都有突出的表现.腺相关病毒(adenoassociated virus,AAV)是一种细小病毒,由于其无致病性和能有效呈递基因而成为一种非常理想的基因表达载体.但是AAV筛除存在一些不足,如对某些重要靶细胞感染效率不高、存在机体的免疫反应等限制了其临床研究的进展.应用DNA改组技术对AAV的衣壳蛋白进行定向进化,有望解决这些问题.本文重点综述DNA改组在AAV衣壳定向进化上的技术发展和应用,并结合本课题组在AAV衣壳DNA改组上的研究工作展开讨论.DNA shuffling is a high-throughput library construction technology that was first described in 1994. Following decades of development, it now has outstanding performance in the reconstruction of DNA, enzymes, and pharmaceutical proteins. The parvovirus adeno- associated virus （AAV） is an ideal gene expression vector because it is non-pathogenic and has potent gene delivery features. However, limitations such as induction of immune response and recalcitrance of some important target cells to AAV infection impede the progress of its clinical study. Using DNA shuffling technology for directed evolution of the AAV capsid protein is expected to resolve these problems. The present paper reviews the technology development and application of DNA shuffling in AAV capsid directed evolution, and discusses the basis of the experiments in this field.

关 键 词：DNA改组 AAV衣壳蛋白 

分 类 号：R440[医药卫生—诊断学]