Microfluidics-Integrated Photothermal Nanoblade for High-Throughput Large Cargo D

用于高通量大型货物 D 的微流控集成光热纳米刀片

• 批准号: 8225967
• 负责人: Pei-Yu Chiou
• 金额: $ 21.78万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-15 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8225967
• 关键词: Age; Analytical Chemistry; Bacteria; Biocompatible Materials; Bioinformatics; Biological; Biomedical Engineering; Cell Aging; Cell Nucleus; Cell Survival; Cell membrane; Cell physiology; Cells; Chromosome Transfer; Chromosomes; Custom; Data; Devices; Disease model; DsRed; Engineering; Epigenetic Process; Event; Explosion; Fibroblasts; Figs - dietary; Film; Frequencies; Goals; Hela Cells; Human; Human Chromosomes; Human Engineering; Investigation; Lasers; Life; Lipofectamine; Liquid substance; Lung; Mammalian Cell; Methods; Microfluidics; Mitochondria; Molecular; Nature; Nuclear Envelope; Organelles; Paper; Pathologic; Physiologic pulse; Physiological; Pluripotent Stem Cells; RNA; Research Infrastructure; Resistance; Somatic Cell; Staging; Subfamily lentivirinae; System; Techniques; Technology; Technology Transfer; Therapeutic; Touch sensation; Work; cell age; cell type; genetic analysis; human embryonic stem cell; human stem cells; inhibitor/antagonist; next generation; novel; pathogen; plasmonics; prototype; success; user-friendly; vapor

DESCRIPTION (provided by applicant): Biomaterial transfer is used in all biological fields to manipulate cell function and dissect molecular and cellular mechanisms in physiologic and pathologic settings. New, enabling technologies are required to overcome size limitations inherent in all current delivery methods to allow the transfer of large cargo, such as live pathogens, whole chromosomes, and (eventually) bioengineered replacement components, such as genetically modified mitochondria, for new-age therapies. As Nicole Rusk, senior editor Nature Methods, recently opined "surprisingly, no methods for efficiently bringing impermeant large molecules into living cells exist (Nat Methods 8:44, 2011). This proposal is focused on our recently developed cargo transfer technology, which is called the photothermal nanoblade, to deliver whole human chromosomes into human pluripotent stem cells (hPSCs) in order to examine chromosome reprogramming. These transferred chromosomes will be analyzed for epigenetic reprogramming within the hPSC nucleus using standard and cutting-edge molecular techniques and is supported by a strong institutional bioinformatics infrastructure. This proposal also aims to develop a microfluidics-integrated photothermal nanoblade platform for next-generation high- throughput and near simultaneous delivery of large cargo into more than 10,000 cells in seconds. Achieving these goals will provide custom engineered hPSCs for biological investigations, an assessment of the chromosome reprogramming capability of federally-registered H1 and H9 human embryonic stem cells (hESCs), and an advanced yet simple to use platform to overcome potential biological obstacles that, if they exist, would limit success due to an insufficient throughput, should chromosome integration be a low frequency event, which is currently not known. PUBLIC HEALTH RELEVANCE: Biomaterial transfer is used in all biological fields to manipulate cell function and dissect molecular and cellular mechanisms in physiologic and pathologic settings. New, enabling technologies are required to overcome size limitations inherent in all current delivery methods to allow the transfer of large cargo for new-age therapies. Our R21 proposal is to use a novel photothermal nanoblade system (Wu, et al., in press, Analytical Chemistry, 2011- see paper in appendix) to deliver whole human chromosomes into pluripotent human stem cells, a currently impossible task, in order to examine whole chromosome epigenetic reprogramming.

描述（由申请人提供）：生物材料转移用于所有生物领域，以操纵细胞功能并剖析生理和病理环境中的分子和细胞机制。需要新的、可行的技术来克服所有当前递送方法固有的尺寸限制，以允许在新时代转移大件货物，例如活病原体、整个染色体，以及（最终）生物工程替代成分，例如转基因线粒体。疗法。正如《自然方法》高级编辑 Nicole Rusk 最近所指出的，“令人惊讶的是，不存在有效地将不渗透性大分子带入活细胞的方法（NatMethods 8:44, 2011）。该提案的重点是我们最近开发的货物转移技术，该技术是称为光热纳米刀片，可将整个人类染色体输送到人类多能干细胞（hPSC）中，以检查染色体重编程，并对这些转移的染色体进行表观遗传分析。该提案还旨在开发一种微流体集成光热纳米刀片平台，用于下一代高通量和近乎同步的大型货物输送​​。在几秒钟内将其转化为超过 10,000 个细胞，将为生物学研究提供定制的 hPSC，以评估联邦注册的 H1 和 H9 人类的染色体重编程能力。胚胎干细胞（hESC），以及一个先进但易于使用的平台，用于克服潜在的生物障碍，如果存在这些障碍，如果染色体整合是一个低频事件（目前尚不清楚），则会因通量不足而限制成功。 公共健康相关性：生物材料转移用于所有生物领域，以操纵细胞功能并剖析生理和病理环境中的分子和细胞机制。需要新的支持技术来克服当前所有输送方法固有的尺寸限制，以允许为新时代疗法转移大量货物。我们的 R21 提案是使用一种新型光热纳米刀片系统（Wu 等人，正在出版，分析化学，2011 年 - 参见附录中的论文）将整个人类染色体传递到多能人类干细胞中，这是目前不可能的任务，以便检查整个染色体表观遗传重编程。