Department of Chemical and Biomolecular Engineering, School of Engineering, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
Beijing Institute of Biotechnology, Beijing, 100071, China.
Adv Mater. 2019 Sep;31(39):e1902469. doi: 10.1002/adma.201902469. Epub 2019 Aug 12.
Cells transport mass dynamically, crossing cell membranes to maintain metabolism and systemic homeostasis, through which biomolecules are also delivered to cells for gene editing, cell reprograming, therapy, and other purposes. Quantifying the translocation kinetics is fundamentally and clinically essential, but remains limited by fluorescence-based technologies, which are semi-quantitative and only provide kinetics information at cellular level or in discrete time. Herein, a real-time method of quantifying cell internalization kinetics is reported using functionalized firefly-luciferase nanocapsules as the probe. This quantitative assay will facilitate the rational design of delivery vectors and enable high-throughput screening of peptides and other functional molecules, constituting an effective tool for broad applications, including drug development and cancer therapy.
细胞通过动态运输物质，穿过细胞膜以维持新陈代谢和全身内稳态，生物分子也通过这种方式被递送到细胞中，用于基因编辑、细胞重编程、治疗等目的。定量研究转运动力学在基础和临床方面都至关重要，但目前仍受到基于荧光的技术的限制，这些技术是半定量的，只能在细胞水平或离散时间提供动力学信息。在此，报道了一种使用功能化萤火虫荧光素酶纳米胶囊作为探针实时定量细胞内化动力学的方法。这种定量分析方法将有助于输送载体的合理设计，并能够对肽和其他功能分子进行高通量筛选，为广泛的应用，包括药物开发和癌症治疗，提供一种有效的工具。
