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Rational design of a genetically encoded infrared fluorescent protease reporter

基因编码红外荧光蛋白酶报告基因的合理设计

基本信息

批准号：
9119028
负责人：
Xiaokun Shu
金额：
$ 31.3万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9119028
关键词：
Animals Apoptosis Apoptotic Binding Biological Biological Process Brain Brain Neoplasms Cardiovascular Diseases Caspase Cells Chemicals Collaborations Cues Development Disease Dorsal Drosophila genus Dyes Embryo Embryonic Development Engineering Fluorescence Resonance Energy Transfer Future Genetic Germ Glioblastoma Glioma Head Health Image Imagery Inhibition of Apoptosis Injection of therapeutic agent Kinetics Lead Malignant Neoplasms Manuscripts Morphogenesis Movement Mus Neurodegenerative Disorders Neurons Oncogene Activation Paper Pattern Peptide Hydrolases Phytochrome Play Property Protein Engineering Proteins Publishing Recruitment Activity Reporter Resolution Role Signal Pathway Signal Transduction Specificity System Time Tissues Transgenic Organisms Work base chromophore cofactor design experience fly image processing improved mouse model spatiotemporal tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Proteases play fundamental roles in almost every major biological process. Changes to proteolytic systems lead to many diseases including cancer, neurodegenerative and cardiovascular diseases. Several chemical dye-based protease reporters have been successful in imaging protease activity in animals. However, these reporters have limitations in imaging developing embryos since it is difficult to target chemical dyes to specific tissues and injection of chemical dyes may perturb development. A genetically encoded protease reporter is thus preferred. Furthermore, compared to fluorescence resonance energy transfer (FRET)-based reporters that suffers from weak signals and require image processing to obtain FRET signal, a fluorogenic protease reporter that becomes fluorescent upon protease activation will be ideal. Such genetically encoded fluorogenic protease reporters will enable us to directly visualize protease activity with spatiotemporal resolution in intact animals. Here we aim to: 1) Design and improve genetically encoded infrared fluorescent protease reporters. Such genetically encoded protease reporters become fluorescent upon protease activation and require no exogenous cofactor. 2): To visualize spatiotemporal dynamics of apoptosis during embryonic morphogenesis. Apoptosis plays an essential role in embryogenesis of Drosophila. Inhibition of apoptosis impairs many critical morphogenetic movements during embryo development, including segmentation, germ band retraction, dorsal closure and head involution. However, it is not clear whether apoptosis is spatiotemporally correlated to morphogenesis. Here, we will use a caspase reporter to visualize spatiotemporal dynamics of apoptosis during these morphogenetic movements. 3): To characterize dynamics of apoptosis during brain tumor development in Drosophila and mice. During cancer development, oncogene activation leads to cell overprolieration, which triggers apoptosis. Evading apoptosis clears this barrier for rapid tumor development, and is one of the hallmarks of cancer. Such dynamics of apoptosis during cancer development has previously been proposed. Here we will use the caspase reporter to characterize dynamics of apoptosis during glioblastoma development in Drosophila and mice.