Channelrhodopsin-Calcium Channel Complexes for Ultrasensitive Optogenetics
用于超灵敏光遗传学的视紫红质通道-钙通道复合物
基本信息
- 批准号:8510730
- 负责人:
- 金额:$ 18.24万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2012
- 资助国家:美国
- 起止时间:2012-07-16 至 2015-01-30
- 项目状态:已结题
- 来源:
- 关键词:AlgaeAnabolismAnimal ModelAppearanceBiological AssayBiomedical ResearchBrainCalcium ChannelCandidate Disease GeneCationsCell membraneCell physiologyCellsChlamydomonas reinhardtiiClinicalClinical TrialsComplementary DNAComplexCoupledDataDrosophila genusFigs - dietaryGene Expression ProfileGene SilencingGenetic TechniquesHealthHumanImmune responseInvestigational TherapiesIon ChannelIonsLightMammalian CellMapsMastigophoraMediatingMembraneMembrane PotentialsMembrane ProteinsMicroRNAsMolecularMonitorMusNeuronsOpticsPhotonsPhotophobiaPhotosensitivityPhototransductionProceduresProcessProteinsResearchResolutionRiskStimulusStressSystemTRP channelTechnologyTestingTherapeuticTherapeutic UsesTimeVariantVisionbaseblindbrain tissuegenome-widelight gatedlight intensitymillisecondnervous system disordernew technologyoptogeneticsoverexpressionpatch clampprotein protein interactionreceptorrestorationsensortool
项目摘要
DESCRIPTION (provided by applicant): Algal channelrhodopsins are light-gated channels widely used for targeted photocontrol of neuron activity. Over the past six years, the temporal and spatial precision of their light-activation has proven to be incisive and powerful for research
on brain circuitry, and more recently channelrhodopsin experimental therapy in animal models of neurological diseases has produced promising results. However, the extremely low ion conductance of channelrhodopsins limits their use and is a significant barrier for human clinical trials because it necessitates high level expression and high light intensities. Overexpression of a foreign membrane protein is detrimental to long-term cellular health and also creates an immune response risk. In stark contrast to the poor photosensitivity in neurons, channelrhodopsins in their native algal cells are low abundance photosensory receptors that mediate phototaxis in extremely low light with near single-photon sensitivity. This exquisite photosensitivity derives from the ~103-fold amplification of channelrhodopsin currents by secondary activation of highly conductive Ca2+ channels in the algal plasma membrane. We propose to develop a new class of ultrasensitive optogenetic tools with the ~103-fold greater light sensitivity by amplification of channelrhodopsin action by co-expression with the Ca2+ channels that naturally perform this function in the alga Chlamydomonas reinhardtii. Our approach is to identify molecular components responsible for amplification of channelrhodopsin-mediated photocurrents in the algae and to construct a combined channelrhodopsin-Ca2+ channel tool. We will apply genome-wide transcriptome profiling coupled to electrophysiological assay of the amplification process to identify the Ca2+ channels and other components necessary for amplification, if they exist. Our evidence strongly favors a direct interaction mechanism, so we expect to identify one or more Ca2+ channels capable of direct amplification by protein-protein interaction. We will further analyze their action by microRNA silencing, and use this information to re-create in mammalian cells an ultrasensitive channelrhodopsin-Ca2+ channel complex used by algae for dim-light phototaxis. We expect this new optogenetic tool to be able to be effective when expressed in the plasma membrane at protein levels well below those that stress human cells. These ultrasensitive molecular complexes will remove a significant barrier to their clinical use, as well as enhance research on neurocircuitry, enabling brain studies not possible with current technology.
描述(由申请人提供):藻类通道视紫红质是广泛用于神经元活性的靶向光控制的光门控通道。在过去的六年里,他们的光激活的时间和空间精度已被证明是精辟和强大的研究
最近,在神经系统疾病的动物模型中进行的通道视紫红质实验疗法已经产生了有希望的结果。然而,通道视紫红质极低的离子电导限制了它们的使用,并且是人类临床试验的重要障碍,因为它需要高水平表达和高光强度。外源膜蛋白的过度表达对长期细胞健康有害,也会产生免疫反应风险。与神经元的光敏性差形成鲜明对比的是,天然藻类细胞中的通道视紫红质是低丰度的感光受体,其在极低的光下以接近单光子的灵敏度介导趋光性。这种精致的光敏性来自于藻细胞质膜中高传导性Ca 2+通道的二次激活导致通道视紫红质电流放大约103倍。我们建议开发一类新的超灵敏的光遗传学工具,通过与天然在莱茵衣藻中执行此功能的Ca 2+通道共表达来放大通道视紫红质作用,从而具有约103倍的光灵敏度。我们的方法是确定负责放大的通道视紫红质介导的光电流在藻类的分子组成部分,并构建一个组合的通道视紫红质-Ca 2+通道工具。我们将应用全基因组转录组分析加上电生理检测的扩增过程中,以确定扩增所需的Ca 2+通道和其他组件,如果它们存在。我们的证据强烈支持直接相互作用机制,因此我们希望确定一个或多个能够通过蛋白质-蛋白质相互作用直接扩增的Ca 2+通道。我们将通过microRNA沉默进一步分析它们的作用,并利用这些信息在哺乳动物细胞中重建藻类用于昏暗光趋光性的超敏感通道视紫红质-Ca 2+通道复合物。我们希望这种新的光遗传学工具能够在质膜中以远低于人类细胞应激水平的蛋白质水平表达时有效。这些超灵敏的分子复合物将消除其临床应用的重大障碍,并加强对神经回路的研究,使目前技术无法进行的大脑研究成为可能。
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(1)
NEUROSCIENCE. Natural light-gated anion channels: A family of microbial rhodopsins for advanced optogenetics.
- DOI:10.1126/science.aaa7484
- 发表时间:2015-08-07
- 期刊:
- 影响因子:0
- 作者:Govorunova EG;Sineshchekov OA;Janz R;Liu X;Spudich JL
- 通讯作者:Spudich JL
Mechanism divergence in microbial rhodopsins.
微生物视紫红质的机制分歧。
- DOI:10.1016/j.bbabio.2013.06.006
- 发表时间:2014
- 期刊:
- 影响因子:0
- 作者:Spudich,JohnL;Sineshchekov,OlegA;Govorunova,ElenaG
- 通讯作者:Govorunova,ElenaG
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
JOHN LEE SPUDICH其他文献
JOHN LEE SPUDICH的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('JOHN LEE SPUDICH', 18)}}的其他基金
Developing an Optogenetics Technology Based on Natural Potassium-selective Channelrhodopsins
开发基于天然钾选择性通道视紫红质的光遗传学技术
- 批准号:
10731153 - 财政年份:2023
- 资助金额:
$ 18.24万 - 项目类别:
Structure/Function of Channelrhodopsins and Related Retinylidene Proteins
视紫红质通道蛋白和相关视黄基蛋白的结构/功能
- 批准号:
10166003 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
Structure/Function of Channelrhodopsins and Related Retinylidene Proteins
视紫红质通道蛋白和相关视黄基蛋白的结构/功能
- 批准号:
10380871 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
Structure/Function of Channelrhodopsins and Related Retinylidene Proteins
视紫红质通道蛋白和相关视黄基蛋白的结构/功能
- 批准号:
10576389 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
Molecular Engineering of Natural Light-Gated Chloride Channels for Optogenetic Inhibition
用于光遗传学抑制的天然光门控氯离子通道的分子工程
- 批准号:
10237959 - 财政年份:2020
- 资助金额:
$ 18.24万 - 项目类别:
Molecular Engineering of Natural Light-Gated Chloride Channels for Optogenetic Inhibition
用于光遗传学抑制的天然光门控氯离子通道的分子工程
- 批准号:
10413162 - 财政年份:2020
- 资助金额:
$ 18.24万 - 项目类别:
Molecular Engineering of Natural Light-Gated Chloride Channels for Optogenetic Inhibition
用于光遗传学抑制的天然光门控氯离子通道的分子工程
- 批准号:
10677649 - 财政年份:2020
- 资助金额:
$ 18.24万 - 项目类别:
Channelrhodopsin-Calcium Channel Complexes for Ultrasensitive Optogenetics
用于超灵敏光遗传学的视紫红质通道-钙通道复合物
- 批准号:
8359246 - 财政年份:2012
- 资助金额:
$ 18.24万 - 项目类别:
Advanced Naturally Designed Channelrhodopsins for Photocontrol of Neural Activity
用于神经活动光控制的先进自然设计通道视紫红质
- 批准号:
7817521 - 财政年份:2009
- 资助金额:
$ 18.24万 - 项目类别:
相似海外基金
Bone-Adipose Interactions During Skeletal Anabolism
骨骼合成代谢过程中骨-脂肪相互作用
- 批准号:
10590611 - 财政年份:2022
- 资助金额:
$ 18.24万 - 项目类别:
Bone-Adipose Interactions During Skeletal Anabolism
骨骼合成代谢过程中的骨-脂肪相互作用
- 批准号:
10706006 - 财政年份:2022
- 资助金额:
$ 18.24万 - 项目类别:
Bone-Adipose Interactions During Skeletal Anabolism
骨骼合成代谢过程中骨-脂肪相互作用
- 批准号:
10368975 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
BCCMA: Foundational Research to Act Upon and Resist Conditions Unfavorable to Bone (FRACTURE CURB): Combined long-acting PTH and calcimimetics actions on skeletal anabolism
BCCMA:针对和抵抗不利于骨骼的条件的基础研究(遏制骨折):长效 PTH 和拟钙剂联合作用对骨骼合成代谢的作用
- 批准号:
10365254 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
Bone-Adipose Interactions During Skeletal Anabolism
骨骼合成代谢过程中骨-脂肪相互作用
- 批准号:
10202896 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
BCCMA: Foundational Research to Act Upon and Resist Conditions Unfavorable to Bone (FRACTURE CURB): Combined long-acting PTH and calcimimetics actions on skeletal anabolism
BCCMA:针对和抵抗不利于骨骼的条件的基础研究(遏制骨折):长效 PTH 和拟钙剂联合作用对骨骼合成代谢的作用
- 批准号:
10531570 - 财政年份:2021
- 资助金额:
$ 18.24万 - 项目类别:
Dissecting molecular mechanisms implicated in age- and osteoarthritis-related decline in anabolism in articular cartilage
剖析与年龄和骨关节炎相关的关节软骨合成代谢下降有关的分子机制
- 批准号:
10541847 - 财政年份:2019
- 资助金额:
$ 18.24万 - 项目类别:
Dissecting molecular mechanisms implicated in age- and osteoarthritis-related decline in anabolism in articular cartilage
剖析与年龄和骨关节炎相关的关节软骨合成代谢下降有关的分子机制
- 批准号:
10319573 - 财政年份:2019
- 资助金额:
$ 18.24万 - 项目类别:
Dissecting molecular mechanisms implicated in age- and osteoarthritis-related decline in anabolism in articular cartilage
剖析与年龄和骨关节炎相关的关节软骨合成代谢下降有关的分子机制
- 批准号:
10062790 - 财政年份:2019
- 资助金额:
$ 18.24万 - 项目类别:
Promotion of NAD+ anabolism to promote lifespan
促进NAD合成代谢以延长寿命
- 批准号:
DE170100628 - 财政年份:2017
- 资助金额:
$ 18.24万 - 项目类别:
Discovery Early Career Researcher Award