Structure-function relationships of AAV capsids in post entry trafficking ans gene transfer

AAV 衣壳在进入后贩运和基因转移中的结构-功能关系

• 批准号: 22731678
• 负责人: Professor Dr. Jürgen Kleinschmidt
• 金额: --
• 依托单位: Abteilung Tumorvirologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/22731678?language=en
• 关键词: Structure; function; relationships; AAV; capsids

Recombinant adeno-associated virus (rAAV) mediated gene transfer is limited at the cellular level by three major barriers: (a) by cell binding and entry, (b) by post entry trafficking and uncoating and (c) by single-strand (ss) to doublestrand (ds) genome conversion. This grant application focuses on the role of a conformational change in the AAV capsid which occurs post entry and is required for viral uncoating and gene transduction. The specific conformational change is defined by the exposure of hidden capsid protein (VP1/VP2) N-termini through pores at the five-fold symmetry axes of the capsid. This in turn leads to the activation of a viral phospholipase activity and possibly further functional elements located on the VP1/VP2 N-termini. The project aims (1) to describe the time point when and in which cellular compartment the conformational change occurs in vivo, (2) to identify the physiological trigger that induces the conformational change, (3) to identify the consequences of VP1/VP2 exposure for AAV post entry trafficking and uncoating, (4) to describe the implications of these post entry events for the development of targeted vectors and (5) to determine if short basic sequence elements located on the VP1/2 N-termini are required for nuclear uptake of the virus.

重组腺相关病毒（rAAV）介导的基因转移在细胞水平上受到三个主要障碍的限制：（a）通过细胞结合和进入，（B）通过进入后运输和脱壳，以及（c）通过单链（ss）到双链（ds）基因组转换。该授权申请集中于AAV衣壳中的构象变化的作用，其发生在进入后并且是病毒脱壳和基因转导所需的。通过在衣壳的五重对称轴处的孔暴露隐藏的衣壳蛋白（VP 1/VP 2）N-末端来定义特定的构象变化。这又导致病毒磷脂酶活性的活化和可能的位于VP 1/VP 2 N-末端上的其他功能元件的活化。该项目旨在（1）描述体内发生构象变化的时间点和细胞区室，（2）鉴定诱导构象变化的生理触发因素，（3）鉴定VP 1/VP 2暴露对AAV进入后运输和脱壳的影响，（4）描述这些进入后事件对于靶向载体的开发的意义，以及（5）确定位于VP 1/2 N-端的短基本序列元件是否与靶向载体的靶病毒的核摄取需要末端。