Latent Luciferase

潜伏荧光素酶

基本信息

批准号：
9193082
负责人：
STEPHEN C. MILLER
金额：
$ 20.94万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
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：
2015-12-15 至 2018-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9193082
关键词：
Acyl Coenzyme A Animal Model Appearance Applications Grants Arachidonic Acids Bacteria Binding Biological Assay Bioluminescence Carboxylic Acids Cells Codon Nucleotides Coenzyme A Ligases Detection Directed Molecular Evolution Drosophila genus Enzymes Evolution Exploratory/Developmental Grant Fireflies Firefly Luciferases Fluorescence Gene Expression Profiling Geometry Image In Vitro Incubated Insecta Ligands Ligase Light Luciferases Mammalian Cell Mammals Methods Modification Mus Mutation Nematoda Order Coleoptera Organism Oxides Plants Process Proteins Reporter Reporting Role Sequence Homology Techniques Testing Therapeutic Visceral Work adenylate analog bioluminescence imaging design drug discovery genetic manipulation in vivo light emission luciferin non-invasive optical imaging oxidation pre-clinical protein function protein protein interaction public health relevance response screening sensor small molecule

项目摘要

DESCRIPTION (provided by applicant): Bioluminescence is a powerful, inexpensive, and non-invasive method to monitor gene expression, enzymatic activity, protein-protein interactions and response to therapeutics both in vitro and in vivo. However, bioluminescence requires the use of a luciferase derived from a small number of luminogenic organisms (e.g., fireflies), and is limited to applications and organisms where genetic manipulation is both possible and practical. In this exploratory/developmental grant, we will identify proteins in non-luminous species that are latent luciferases, capable of bioluminescent light emission when treated with an appropriate substrate. In support of this substrate-directed approach, we have identified a fruit fly protein that is revealed to be a luciferase when treated with a synthetic luciferin analog. This unprecedented result shows that the function of a protein can be fundamentally changed simply by modification of the substrate, rather than by mutation. Beyond the paradigm-shifting implications for the evolution and design of new enzymatic activities, the prospect that non-luminogenic organisms encode latent luciferases has transformative potential for the non-invasive optical imaging of endogenous processes, potentially enabling bioluminescence imaging in native cells and organisms without the need for the introduction of an exogenous luciferase.

描述（由申请人提供）：生物发光是一种强大、廉价且非侵入性的方法，用于监测体外和体内的基因表达、酶活性、蛋白质-蛋白质相互作用以及对治疗的反应。荧光素酶源自少数发光生物（例如萤火虫），并且数量有限在这项探索性/开发性资助中，我们将鉴定出非发光物种中的蛋白质，这些蛋白质是潜在的荧光素酶，当用适当的底物处理时能够发出生物发光，以支持这一点。通过底物导向方法，我们发现了一种果蝇蛋白，当用合成的荧光素类似物处理时，该蛋白被证明是一种荧光素酶，这一前所未有的结果表明，只需对蛋白进行修饰，就可以从根本上改变蛋白质的功能。除了对新酶活性的进化和设计产生范式转变的影响之外，非发光生物编码潜在荧光素酶的前景对于内源过程的非侵入性光学成像具有变革潜力，有可能实现生物发光。在天然细胞和生物体中成像，无需引入外源荧光素酶。