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: 10661807
• 负责人: Naihao Chiang
• 金额: $ 24.65万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661807
• 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; Malignant Neoplasms; Membrane; Methods; Microfluidics; Microscope; Modality; Monitor; Optics; Penetration; Performance; Phase; Physics; Population; Positioning Attribute; Process; Public Health; Regenerative Medicine; Research; Route; Scanning; Scheme; Site; Spectrum Analysis; Surface Plasmon Resonance; System; Technology; Toxic effect; Transfection; Work; cell type; clinical implementation; delivery vehicle; design; embryonic stem cell; gene therapy; genome editing; imaging capabilities; imaging platform; improved; innovation; nano; nanoscale; next generation; parallelization; plasmonics; prototype; research and development; scanning ion conductance microscopy; single molecule; stem; stem cell biology; stem cell genes; stem cells; targeted delivery; targeted nucleases; therapeutic genome editing; tool; translational medicine; treatment strategy

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.

摘要 最近出现的强大的基因组编辑方法（CRISPR相关的靶向核酸酶技术） 以及它们在干细胞中的应用，导致了研究和开发的革命性突破， 下一代基因编辑疗法为了便于转染，需要将基因编辑材料 以快速、有效和安全的方式递送到细胞中。虽然一些物理和生物化学的 细胞内递送的方法现在在实验室环境中常规使用，但是效率，通量， 和毒性限制了它们的临床应用， 细胞 为了直接应对上述挑战，该提案旨在开发一种高精度的 等离子体诱导的细胞内递送方案，其利用高度局部化和增强的 在等离子纳米移液管附近的电磁场。目标是开发一个综合的 平台，重点是干细胞基因编辑，用于细胞内递送的通用方法， 所有种类的货物和细胞类型的表征具有高效率和活力，从单细胞到 数百万个细胞。将开发两个原型平台：i）用于非接触式等离子体纳米移液管， 通过与扫描离子电导显微镜结合的细胞内递送;和ii） 等离子纳米移液管通过惯性微流体。成功地实现这两个目标将导致 - 实现通用的细胞内递送而不物理穿透细胞膜， 单货物单细胞精确度此外，近场光学传感将被引入到开发的 平台它将为单分子非侵入性、连续、无标记的生物传感提供一条直接途径 灵敏度因此，不仅等离子体诱导的递送的基本机制将被询问， 但它也将提供现场反馈，以进一步改进已开发的平台。

项目成果

Large-Scale Soft-Lithographic Patterning of Plasmonic Nanoparticles.

• DOI: 10.1021/acsmaterialslett.0c00535
• 发表时间: 2021-03-01
• 期刊: ACS materials letters
• 影响因子: 11.4
• 作者: Chiang N;Scarabelli L;Vinnacombe-Willson GA;Pérez LA;Dore C;Mihi A;Jonas SJ;Weiss PS
• 通讯作者: Weiss PS

Seeded-Growth Experiment Demonstrating Size- and Shape-Dependence on Gold Nanoparticle-Light Interactions.

• DOI: 10.1021/acs.jchemed.0c01150
• 发表时间: 2021-02-09
• 期刊: Journal of chemical education
• 影响因子: 3
• 作者: Vinnacombe-Willson GA;Chiang N;Weiss PS;Tolbert SH;Scarabelli L
• 通讯作者: Scarabelli L

In Situ Shape Control of Thermoplasmonic Gold Nanostars on Oxide Substrates for Hyperthermia-Mediated Cell Detachment.

• DOI: 10.1021/acscentsci.0c01097
• 发表时间: 2020-11-25
• 期刊: ACS central science
• 影响因子: 18.2
• 作者: Vinnacombe-Willson GA;Chiang N;Scarabelli L;Hu Y;Heidenreich LK;Li X;Gong Y;Inouye DT;Fisher TS;Weiss PS;Jonas SJ
• 通讯作者: Jonas SJ