Synthetic adenovirus libraries for vector optimization

用于载体优化的合成腺病毒文库 腺病毒已被广泛用于解决生物医学研究中的基本问题。这些 小的dna病毒依赖于大量的宿主蛋白进行自身复制，并一直是有用的探针。 探索哺乳动物细胞的基本工作原理。基因上易处理的重组腺病毒 已被广泛用作基因传递的载体。理性的和不偏不倚的方法都是 用来增强和修改它们的功能特性。向量优化的合理途径 受到我们对许多病毒功能的不完全理解的限制，而公正的方法是 技术上受到基因组大小和诱变特定基因的固有困难的限制 利息。这项建议的重点是开发一个新颖的平台，用于创建定制的 腺病毒文库。该项目的关键创新是最近开发的用于智能网的系统 腺病毒基因组的体外组装来自紧凑的模块，可以单独操作，然后 重新组装。在这个项目中，将使用合成方法来生成高多样性的焦点区域 在整个人类基因组中，腺病毒5是使用最广泛的载体血清型。由此产生的DNA 子库将被组装成完整的病毒基因组，每个组装都分配了一个唯一的条形码 旨在促进文库表征和个体突变体的跟踪。建筑和建筑 高含量、局灶腺病毒文库的特征及其用途将通过一个简单的 筛选可逃避中和单抗识别的变异病毒。的长期目标是 这个项目是开发一个资源，可以共享和不断发展，以满足不断发展的需求 不同学科的科学家。

Synthetic adenovirus libraries for vector optimization Adenoviruses have been extensively utilized to address fundamental questions in biomedical research. These small DNA viruses rely on numerous host proteins for their own replication, and have been useful probes to explore the fundamental workings of the mammalian cell. Genetically tractable, recombinant adenoviruses have become widely used as vectors for gene delivery. Both rational and unbiased methods have been employed to enhance and modify their functional characteristics. Rational approaches to vector optimization are constrained by our incomplete understanding of many viral functions, while unbiased methods are technically limited by the size of the genome and the inherent difficulty of mutagenizing specific genes of interest. This proposal is focused on the development of a novel platform for the creation of customized adenovirus libraries. The critical innovation that underlies this project is a recently developed system for the in vitro assembly of adenovirus genomes from compact modules that can be individually manipulated and then reassembled. In this project, synthetic methods will be employed to generate focal regions of high diversity across the genome of human adenovirus 5, the most widely-employed vector serotype. The resulting DNA sub-libraries will be assembled into complete viral genomes, with each assembly assigned a unique barcode designed to facilitate library characterization and the tracking of individual mutants. The construction and characterization of high-content, focal adenoviral libraries, and their utility, will be demonstrated by a simple screen for mutant viruses that evade recognition by a neutralizing monoclonal antibody. The long term goal of this project is to develop a resource that can be shared and continually developed to meet the evolving needs of scientists across many disciplines.