Transgenic Tethered Peptides

描述（由申请人提供）：我们的长期目标是开发工具，用于表征神经回路和非神经元组织在完整行为动物体内的功能。我们已经开发了一种新技术，将神经肽作为嵌合融合蛋白通过疏水锚定（“t肽”）拴在质膜上进行转基因表达。t肽在表达t肽的细胞中以预期的特异性激活其同源g蛋白偶联受体（gpcr），但不激活邻近不表达t肽的细胞膜中的gpcr。因此，T肽为转基因动物神经肽功能的细胞自主获得分析提供了遗传编码工具。特异性目标旨在提供一个全基因组工具包的基因编码细胞自主药理学特异性激活的神经肽gpcr。由于该工具包将基于UAS-GAL4二元表达系统，该系统将细胞和电路特异性启动子GAL4驱动基因从含有编码效应（如t肽）的cdna的UAS转基因中分离出来，因此它将直接用于对神经肽的细胞特异性生理和行为功能感兴趣的果蝇生物学家。更一般地说，t肽文库的开发和验证也将提供宝贵的见解，使能够产生用于研究哺乳动物神经肽在健康和疾病中的体内功能的工具，包括基因治疗应用的潜在临床应用。

DESCRIPTION (provided by applicant): Our long-term goal is the development of tools for characterizing the functions of neuropeptides in neural circuits and non-neuronal tissues in vivo in intact behaving animals. We have developed a new technology, where neuropeptides are transgenically expressed as chimeric fusion proteins tethered to the plasma membrane via hydrophobic anchors ("t-peptides"). T-peptides activate their cognate G-protein coupled receptors (GPCRs) with expected specificity in t-peptide-expressing cells, but without activating their GPCRs in the membranes of neighboring cells not expressing the t-peptide. T- peptides thus provide genetically encoded tools for the cell-autonomous gain-of-function analysis of neuropeptide function in transgenic animals. The Specific Aims are designed to provide a genome-wide toolkit of genetically encoded cell-autonomous pharmacologically specific activators of neuropeptide GPCRs. Because this toolkit will be based on the UAS-GAL4 binary expression system that separates cell- and circuit- specific promoter GAL4 driver transgenes from UAS transgenes containing cDNAs that encode effectors such as t-peptides, it will be of direct utility to Drosophila biologists interested in cell-specific physiological and behavioral functions of neuropeptides. More generally, the development and validation of t-peptide libraries will also provide invaluable insights enabling the generation of tools for use in addressing mammalian neuropeptide function in vivo in health and disease, including potential clinical utility in gene therapy applications.