Rewiring Cell Membranes with Bioorthogonal Surface Chemistry for Fundamental and Applied Studies

在空间和时间上引导由多个细胞系组成的三维（3-D）组织组装的能力对于免疫和再生医学中的下一代治疗应用至关重要。高效的核酸转染、体内成像、追踪和靶向药物递送至特定细胞、细胞组织和耐药细菌对于许多诊断生物技术、疫苗开发和药物筛选平台至关重要。这些研究领域和市场是多样化的，但集中在为特定应用重新布线（工程）细胞表面的能力。一种新的通用方法，重新布线细胞表面的能力，安装任何配体或蛋白质的细胞表面上，也能够控制细胞在空间和时间上的相互连接，将允许前所未有的探索一系列的基本细胞行为的研究，并提供新的方式来显示成像探针，推进细胞为基础的生物技术和加速再生医学和组织工程为基础的治疗。在本建议中，我们将制定一个强大的通用战略，即：1。通过纳米技术（脂质体融合）将一系列生物正交化学基团递送到细胞表面，用于随后的原位定制，用于一系列新的生物技术、转染和组织工程应用的按需组织组装，以及2.在不使用分子生物学或代谢生物合成的情况下将广泛的配体或蛋白质快速递送至细胞表面;以及3.一种结合细胞表面工程和生物正交化学的方法，通过点击化学用核酸转染细胞。

The ability to spatially and temporally direct 3-dimensional (3-D) tissue assembly comprised of multiple cell lines is crucial for next generation therapeutic applications in immunity and regenerative medicine. Efficient nucleic acid transfection, In vivo Imaging, tracking and targeted drug delivery to specific cells, cell tissues and drug resistant bacteria is critical for many diagnostic biotechnologies, vaccine development and drug screening platforms. These research fields and markets are diverse but converge at the ability to rewire (engineer) cell surfaces for specific applications. A new general method that rewires cell surfaces with the capability to install any ligand or protein on a cell surface and to also be able to control cell to cell interconnectivity in space and time would allow for unprecedented exploration of a range of fundamental cell behaviour studies and to provide new ways to display imaging probes, advance cell based biotechnologies and accelerate regenerative medicine and tissue engineering based therapies. In this proposal, we will develop a powerful universal strategy that: 1. delivers a range of bio-orthogonal chemical groups via nanotechnology (liposome fusion) to cell surfaces for subsequent in-situ tailoring for on-demand tissue assembly for a range of new biotechnology, transfection and tissue engineering applications and 2. delivers a broad range of ligands or proteins rapidly to cell surfaces without the use of molecular biology or metabolic biosynthesis and 3. a combined cell surface engineering and bioorthogonal chemistry approach to transfect cells with nucleic acid via click chemistry.