Monitoring expression of cell associated transgenes /vector core

该项目/核心的目标是：1）开发一种方便、可靠和普遍适用的策略，以监测体内任何载体编码转基因的表达; 2）为所有PPG研究者提供根据其个人需求定制的逆转录病毒、慢病毒和腺病毒基因治疗载体的充分表征制剂。在任何体内基因治疗研究中，治疗性转基因表达谱是结果的关键决定因素，但通常无法获得此信息。本项目的总体假设是，治疗性转基因的表达可以通过测量从相同载体构建体一致表达的可溶性、非免疫原性标记多肽在体液中的浓度而在体内非侵入性地监测。具体目标是：1）精确定义载体编码的多肽的合成速率之间的一致性，所述多肽的表达通过内部核糖体进入位点（IRES）连接; 2）为了确定体内基因表达谱和从逆转录病毒制剂的双顺反子慢病毒和腺病毒载体表达的可溶性标记肽的体液浓度之间随时间的一致性，慢病毒或腺病毒载体，其结合了任选的一致表达的β-hCG标记多肽的特征，以支持它们在免疫活性大鼠、狗和猪中的临床前研究。

The goals of this project/core are: 1) to develop a convenient , reliable and generally applicable strategy to monitor in vivo the expression of any vector encoded transgene; and 2) to supply all PPG investigators with well-characterized preparations of retroviral, lentiviral and adenoviral gene therapy vectors tailored to their individual requirements. The profile of therapeutic transgene expression is a key determinant of the outcome in any in vivo gene therapy study, but this information is usually not available. The overall hypothesis for the current project is that the expression of the therapeutic transgene can be non-invasively monitored in vivo by measuring the concentration in body fluids of a soluble, non- immunogenic marker polypeptide concordantly expressed from the same vector construct. The Specific Aims are: 1) To precisely define the concordance between the rates of synthesis of vector encoded polypeptides whose expression is linked through an internal ribosome entry site (IRES); 2) To define the in vivo gene expression profiles and concordance over time between body fluid concentrations of soluble marker peptides expressed from the bicistronic lentiviral and adenoviral vectors of preparations of retroviral, lentiviral or adenoviral vectors incorporating the optional feature of concordantly expressed beta-hCG marker polypeptide to support their preclinical studies in immunocompetent rats, dogs and pigs.