ANALYSIS OF CALI EXPERIMENTS WITH VIRTUAL CELL
虚拟细胞 CALI 实验分析
基本信息
- 批准号:8169581
- 负责人:
- 金额:$ 1.63万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2010
- 资助国家:美国
- 起止时间:2010-05-01 至 2011-04-30
- 项目状态:已结题
- 来源:
- 关键词:ActinsCell LineCell physiologyCellsChimeric ProteinsComplementComputer Retrieval of Information on Scientific Projects DatabaseDendritic CellsFilamentFundingGenerationsGrantGrowthInstitutionLabelLasersLightMediatingMicrofilamentsModelingPhenotypePhysiologicalProteinsReagentResearchResearch PersonnelResourcesRoleSourceStructureSystemTechniquesUnited States National Institutes of Healthchromophoreknock-downloss of functionresearch studyspatiotemporalvirtual
项目摘要
This subproject is one of many research subprojects utilizing the
resources provided by a Center grant funded by NIH/NCRR. The subproject and
investigator (PI) may have received primary funding from another NIH source,
and thus could be represented in other CRISP entries. The institution listed is
for the Center, which is not necessarily the institution for the investigator.
Chromophore-assisted laser inactivation (CALI) is a light-mediated technique that offers precise spatiotemporal control of protein inactivation, enabling better understanding of the protein's role in cell function. EGFP has been used effectively as a CALI chromophore, and its cotranslational attachment to the target protein avoids having to use exogenously added labeling reagents. A potential drawback to EGFP-CALI is that the CALI phenotype can be obscured by the endogenous, unlabeled protein that is not susceptible to light inactivation. Performing EGFP-CALI experiments in deficient cells rescued with functional EGFP-fusion proteins permits more complete loss of function to be achieved. We developed a modified lentiviral system for rapid and efficient generation of knockdown cell lines complemented with physiological levels of EGFP-fusion proteins. We demonstrated that CALI of EGFP-CapZbeta increases uncapped actin filaments, resulting in enhanced filament growth and the formation of numerous protrusive structures. We showed that these effects are completely dependent upon knocking down the endogenous protein. To fully interpret these results and also guard against over-interpretation, a quantitative analysis, which build on the Virtual Cell dendritic actin nucleation model, will be developed.
这个子项目是众多研究子项目之一
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Kenneth A Jacobson其他文献
Kenneth A Jacobson的其他文献
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{{ truncateString('Kenneth A Jacobson', 18)}}的其他基金
Structure and dynamics of membrane microdomains used for viral entry and egress
用于病毒出入的膜微域的结构和动力学
- 批准号:
7999969 - 财政年份:2010
- 资助金额:
$ 1.63万 - 项目类别:
2004 Biophysical Discussion: Membrane Microdomains
2004 年生物物理讨论:膜微域
- 批准号:
6834500 - 财政年份:2004
- 资助金额:
$ 1.63万 - 项目类别:
REGULATION OF MOTILITY AND TRANSCRIPTION IN INFLAMMATION
炎症中运动和转录的调节
- 批准号:
6654106 - 财政年份:2002
- 资助金额:
$ 1.63万 - 项目类别:
REGULATION OF MOTILITY AND TRANSCRIPTION IN INFLAMMATION
炎症中运动和转录的调节
- 批准号:
6644954 - 财政年份:2001
- 资助金额:
$ 1.63万 - 项目类别:
REGULATION OF MOTILITY AND TRANSCRIPTION IN INFLAMMATION
炎症中运动和转录的调节
- 批准号:
6300943 - 财政年份:1999
- 资助金额:
$ 1.63万 - 项目类别:
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