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Control of Protein Activity Via Light Stimulated Dimerizers

通过光刺激二聚体控制蛋白质活性

基本信息

批准号：
9324407
负责人：
Chandra L Tucker
金额：
$ 23.84万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9324407
关键词：
Ablation Acute Adopted Autophagocytosis Biochemical Process Biological Biological Process Brain Cell physiology Cellular biology Chemicals DNA Developmental Cell Biology Dimerization Disease Dose Engineering Enzymes Event Funding Gene Expression Regulation Gene Proteins Generations Genes Genetic Genetic Recombination Genetic Transcription Goals Health Homodimerization Hour Ion Channel Ion Pumps Knock-out Label Light Location Methods Modeling Molecular Mouse-ear Cress Neurobiology Neurons Optics Pathway interactions Peptide Hydrolases Photoreceptors Phototransduction Physiological Processes Plant Proteins Post-Translational Protein Processing Process Property Protein Fragment Proteins Proteolysis RNA Regulation Research Research Personnel Rodent Role S-nitro-N-acetylpenicillamine Signal Transduction Site System Technology Testing Time Tissue Engineering Ubiquitin Validation Variant Work base chemical genetics cryptochrome cryptochrome 2 genetic approach genetic selection improved in vivo insight knock-down novel optogenetics protein function recombinase reconstitution research study synthetic biology temporal measurement tool transcription factor

项目摘要

This work seeks to engineer, optimize, and validate new tools and strategies to control protein activity with light, which can be delivered with spatial and temporal resolution. In previous work, we pioneered use of the Arabidopsis thaliana blue light photoreceptor cryptochrome 2 (CRY2) as an optogenetic module. We developed a light-induced dimerization platform based on interaction with a partner protein, CIB1, which can be used to control activity and localization of target proteins. In recent work, we developed a new photoregulated module, CRY2olig, that induces protein oligomerization with light. The broad aims of this project are to further develop the CRY2/CIB and CRY2olig technologies allowing control of protein dimerization and oligomerization with light. Specifically, we aim to optimize CRY2/CIB dimerizers with improved and extended properties for cell biology applications (Aim 1). In Aim 2, we will apply these dimerizers to control activity of a set of important and versatile regulatory enzymes, allowing manipulation of DNA recombination, protein cleavage, and protein labeling with light. Importantly, we will carry out further optimization and validation of a promising light-activated Cre recombinase, activity of which we will test in vivo in rodent brain. In Aim 3, we will develop and validate methods to transiently and locally disrupt protein activity using CRY2 and CRY2olig. Such tools will enable researchers to quickly and precisely disrupt function, avoiding compensatory effects that can occur with genetic ablation or knockdown studies.

这项工作旨在设计，优化和验证新的工具和策略来控制蛋白质活性