OptoTFs: Genetically encoded light-activated transcription factors

OptoTFs：基因编码的光激活转录因子

• 批准号: 7830210
• 负责人: SAMIE R JAFFREY
• 金额: $ 16.95万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7830210
• 关键词: Address; Animals; Behavior; CREB1 gene; Cell physiology; Cells; Chimeric Proteins; Cleaved cell; Couples; Dendrites; Dependence; Development; Fiber Optics; Future; Gene Expression; Genetic Transcription; Goals; Lead; Learning; Life; Light; Light Exercise; Mammalian Cell; Memory; Methods; Molecular; Morphogenesis; Neurons; Neurophysiology - biologic function; Neurosciences; Opsin; Pattern; Peptides; Proteins; Proteolysis; Regulation; Role; Slice; Synapses; System; Techniques; Testing; Tissues; Transcriptional Activation; Vertebral column; activating transcription factor; addiction; brain behavior; design; design and construction; drug of abuse; insight; microbial; multicatalytic endopeptidase complex; novel; novel strategies; protein degradation; protein function; prototype; research study; tool; tool development; transcription factor

DESCRIPTION (provided by applicant): The ability to control neuronal function in cells, tissues, and living animals is critical for understanding the role of specific proteins in neural function and behavior. Controlling protein function by light has emerged as a popular approach due to its limited effects on endogenous cellular function. However, other than microbial opsins, few proteins when expressed in mammalian cells can be regulated by light. This application describes the design, construction, optimization, and application of a new class of proteins, the optoTFs, which are genetically encoded, light-activated transcription factors. OptoTFs will allow gene transcription in cells, tissue, or animals to be controlled by light in a spatially and temporally controlled manner. Experiments in this application seek to optimize the construction of an OptoTF that has CREB activity following light stimulation. The ability of light to activate CREB activity in dissociated neuronal culture and in organotypic slice culture will be optimized. Using this optoTF, the role of CREB in dendrite morphogenesis will also be investigated. The experiments in this application will develop a fundamentally novel approach to use light to spatially and temporally manipulate gene transcription. Furthermore, the methods established here are likely to be generalizable to other types of proteins, thereby potentially enabling the optogenetic control of a wide range of proteins for numerous applications. This application describes the design, construction, optimization, and application of a new class of proteins, the optoTFs, which are genetically encoded, light-activated transcription factors. OptoTFs will serve as a novel tool for the control of gene expression in cells, tissues and living animals. The development of this tool will enable future studies to establish the role of gene transcription in learning, memory, and addiction.

描述（由申请人提供）：控制细胞、组织和活体动物中神经元功能的能力对于理解特定蛋白质在神经功能和行为中的作用至关重要。由于光对内源细胞功能的影响有限，因此通过光控制蛋白质功能已成为一种流行的方法。然而，除了微生物视蛋白之外，在哺乳动物细胞中表达的蛋白质很少能受到光的调节。该应用描述了一类新型蛋白质 optoTF 的设计、构建、优化和应用，optoTF 是基因编码的光激活转录因子。 OptoTF 允许光以空间和时间控制的方式控制细胞、组织或动物中的基因转录。本应用中的实验旨在优化 OptoTF 的构建，该 OptoTF 在光刺激后具有 CREB ​​活性。光在分离神经元培养物和器官切片培养物中激活 CREB ​​活性的能力将得到优化。使用该 optoTF，还将研究 CREB ​​在树突形态发生中的作用。该应用中的实验将开发一种全新的方法，利用光在空间和时间上操纵基因转录。此外，这里建立的方法可能适用于其他类型的蛋白质，从而有可能实现对多种蛋白质的光遗传学控制，以用于众多应用。该应用描述了一类新型蛋白质 optoTF 的设计、构建、优化和应用，optoTF 是基因编码的光激活转录因子。 OptoTF 将成为控制细胞、组织和活体动物基因表达的新工具。该工具的开发将使未来的研究能够确定基因转录在学习、记忆和成瘾中的作用。