High-Precision Non-Contact Plasmon-Induced Intracellular Delivery

高精度非接触式等离激元诱导细胞内递送

• 批准号: 9806813
• 负责人: Naihao Chiang
• 金额: $ 8.98万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-05 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9806813
• 关键词: Address; Anemia; Basic Science; Biochemical; Biosensing Techniques; Cell Nucleus; Cell Survival; Cell Therapy; Cell membrane; Cells; Chemistry; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coupled; Custom; Cystic Fibrosis; Devices; Diagnostic; Discipline; Disease; Doctor of Philosophy; Electromagnetic Fields; Electroporation; Environment; Feedback; Gene Delivery; Genes; Goals; Gold; Health; Hour; In Situ; Ion Channel; Ions; Label; Laboratories; Lasers; Lead; Malignant Neoplasms; Membrane; Methods; Microfluidics; Microscope; Modality; Monitor; Optics; Performance; Phase; Physics; Population; Positioning Attribute; Process; Public Health; Regenerative Medicine; Research; Route; Scanning; Scheme; Site; Spectrum Analysis; Stem cells; Surface Plasmon Resonance; System; Technology; Toxic effect; Transfection; Work; base; cell type; clinical implementation; design; embryonic stem cell; gene therapy; genome editing; imaging capabilities; imaging platform; improved; innovation; nanoscale; next generation; parallelization; plasmonics; prototype; research and development; scanning ion conductance microscopy; single molecule; stem; stem cell biology; targeted delivery; targeted nucleases; tool; translational medicine; treatment strategy; vector

ABSTRACT The recent emergence of robust genome editing methods (CRISPR associated targeted nuclease technologies) and their applications to stem cells has led to revolutionary breakthrough in the research and development of next-generation gene-editing therapies. In order to facilitate the transfection, gene-editing materials need to be delivered into cells in a rapid, efficient, and safe manner. Although some of the physical and biochemical methods for intracellular delivery are now used routinely in laboratory settings, issues with efficiency, throughput, and toxicity have limited their clinical implementations for universal delivering all-sizes of cargos into all-types of cells. To directly address the aforementioned challenges, this proposal aims to develop a high-precision plasmon-induced intracellular delivery scheme which utilizes the highly localized and intensified electromagnetic field in the close proximity of the plasmonic nanopipettes. The goal is to develop an integrated platform, with an emphasis on stem cell gene-editing, for universal approach of intracellular delivery and characterization for all varieties of cargo and cell types with high-efficiency and -viability from single-cell to millions of cells. Two prototype platforms will be developed: i) plasmonic nanopipettes for non-contact intracellular delivery through combining with scanning ion conductance microscopy; and ii) Parallelization of plasmonic nanopipettes via inertial microfluidics. Successfully completion of these two aims would lead to realization of universal intracellular delivery without physically penetrating the cell membranes, with single-cargo, single-cell precision. Furthermore, near-field optical sensing will be introduced into the developed platforms. It will provide a direct route to non-invasive, continuous, label-free biosensing with single-molecule sensitivity. Therefore, not only the fundamental mechanism of the plasmon-induced delivery will be interrogated, but it will also provide an in-situ feedback to further improvement of the developed platforms.

抽象的 强大的基因组编辑方法的最新出现（CRISPR相关的靶向核酸酶技术） 它们对干细胞的应用导致了在研究和发展的革命突破 下一代基因编辑疗法。为了促进转染，基因编辑材料必须为 以快速，高效且安全的方式输送到细胞中。虽然有些物理和生化 现在，在实验室环境中常规使用细胞内交付的方法，效率，吞吐量，吞吐量， 毒性将其临床实施限制为通用全尺寸的赤道 细胞。 为了直接应对上述挑战，该提案旨在发展高精度 血浆诱导的细胞内递送方案，利用高度局部和加强 等离激元纳米夹的近距离电磁场。目标是开发一个综合的 平台，重点是干细胞基因编辑，用于通用细胞内递送和 各种货物和细胞类型的特征，具有高效且可视性。 数百万个细胞。将开发两个原型平台：i）非接触的等离子体纳米夹 通过与扫描离子电导显微镜结合的细胞内递送； ii）并行 血浆纳米管通过惯性微流体。成功完成这两个目标将导致 实现通用细胞内递送，而没有物理穿透细胞膜， 单卡，单细胞精度。此外，将引入近场光学传感 平台。它将提供直接途径，直接通往单分子的非侵入性，连续，无标签的生物传感 灵敏度。因此，不仅等离子体引起的输送的基本机制将受到询问，而且还会被审问 但这还将提供现场反馈，以进一步改善开发的平台。