Biomacromolecules Research Team, RIKEN Center for Sustainable Resource Science, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Kyoto-Daigaku-Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Biomacromolecules. 2020 Jul 13;21(7):2735-2744. doi: 10.1021/acs.biomac.0c00481. Epub 2020 Jun 3.
Owing to their diverse functions and tunable physicochemical properties, peptides are promising alternatives to the conventional gene delivery tools that are available for plant systems. However, peptide-mediated gene delivery is limited by low transfection efficiency in plants because of the insufficient cytosolic translocation of DNA cargo. Here, we report a dual peptide-based gene delivery system for the efficient transfection of plant callus cells. This system is based on the combination of an artificial peptide composed of cationic cell-penetrating and hydrophobic endosomal escape domains with a gene carrier peptide composed of amphiphilic cell-penetrating and cationic DNA-binding domains. Cellular internalization and transfection studies revealed that this dual peptide-based system enables more efficient transfection of callus cells than does a carrier peptide alone by enhancing the endocytic uptake and subsequent cytosolic translocation of a carrier peptide/DNA complex. The present strategy will expand the utility of peptide-mediated plant gene delivery for a wide range of applications and basic research.
由于其多样化的功能和可调节的物理化学性质，肽类是传统基因传递工具的有前途的替代品，可用于植物系统。然而，由于 DNA 货物在细胞质中的转运不足，肽介导的基因传递在植物中受到限制。在这里，我们报告了一种基于双肽的基因传递系统，用于有效转染植物愈伤组织细胞。该系统基于阳离子细胞穿透和内体逃逸结构域的人工肽与由两亲性细胞穿透和阳离子 DNA 结合结构域组成的基因载体肽的组合。细胞内化和转染研究表明，与单独的载体肽相比，这种基于双肽的系统通过增强载体肽/DNA 复合物的内吞摄取和随后的细胞质转运，能够更有效地转染愈伤组织细胞。本策略将扩大肽介导的植物基因传递在广泛的应用和基础研究中的效用。
