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

在空间和时间上指导由多个细胞系组成的三维（3-D）组织组装的能力对于下一代免疫和再生医学的治疗应用至关重要。高效的核酸转染、体内成像、跟踪和靶向药物递送到特定细胞、细胞组织和耐药细菌对许多诊断生物技术、疫苗开发和药物筛选平台至关重要。这些研究领域和市场各不相同，但都集中在为特定应用重新布线（工程）细胞表面的能力上。一种新的通用方法可以重新连接细胞表面，能够在细胞表面安装任何配体或蛋白质，并且能够控制细胞与细胞在空间和时间上的相互连接，这将允许对一系列基本细胞行为研究进行前所未有的探索，并提供展示成像探针的新方法，推进基于细胞的生物技术，加速再生医学和组织工程治疗。在这一建议中，我们将制定一个强有力的普遍战略，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输送到细胞表面，用于随后的原位剪裁，用于一系列新的生物技术、转染和组织工程应用。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到细胞表面。结合细胞表面工程和生物正交化学方法，通过点击化学转染核酸细胞。

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.