Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Biomacromolecules Research Team, RIKEN Center for Sustainable Resource Science, 2-1 Hirosawa, Wako, Saitama 3510198, Japan.
Biomacromolecules. 2023 Aug 14;24(8):3657-3665. doi: 10.1021/acs.biomac.3c00391. Epub 2023 Jun 29.
Plant mitochondria play essential roles in metabolism and respiration. Recently, there has been growing interest in mitochondrial transformation for developing crops with commercially valuable traits, such as resistance to environmental stress and shorter fallow periods. Mitochondrial targeting and cell membrane penetration functions are crucial for improving the gene delivery efficiency of mitochondrial transformation. Here, we developed a peptide-based carrier, referred to as Cytcox/KAibA-Mic, that contains multifunctional peptides for efficient transfection into plant mitochondria. We quantified the mitochondrial targeting and cell membrane-penetrating peptide modification rates to control their functions. The modification rates were easily determined from high-performance liquid chromatography chromatograms. Additionally, the gene carrier size remained constant even when the mitochondrial targeting peptide modification rate was altered. Using this gene carrier, we can quantitatively investigate the relationships between various peptide modifications and transfection efficiency and optimize the gene carrier conditions for mitochondrial transfection.
植物线粒体在代谢和呼吸中发挥着重要作用。最近，人们对线粒体转化越来越感兴趣，希望利用这一技术开发具有商业价值的作物，例如具有环境胁迫抗性和缩短休耕期的作物。线粒体靶向和细胞膜穿透功能对于提高线粒体转化的基因传递效率至关重要。在这里，我们开发了一种基于肽的载体，称为 Cytcox/KAibA-Mic，它包含多功能肽，可有效转染植物线粒体。我们量化了线粒体靶向和细胞膜穿透肽的修饰率，以控制其功能。修饰率可以很容易地从高效液相色谱图中确定。此外，即使改变线粒体靶向肽的修饰率，基因载体的大小也保持不变。使用这种基因载体，我们可以定量研究各种肽修饰与转染效率之间的关系，并优化线粒体转染的基因载体条件。
