Gene introduction into the mitochondria of Arabidopsis thaliana via peptide-based carriers.

Gene introduction into the mitochondria of Arabidopsis thaliana via peptide-based carriers.
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DOI:
10.1038/srep07751
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发表时间:
2015-01-13
期刊:
影响因子:
4.6
通讯作者:
Numata K
Numata K
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Chuah JA;Yoshizumi T;Kodama Y;Numata K

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植物遗传转化中可用的方法是核转化和质体转化,因为线粒体的类似程序尚未建立。细胞器的双膜和小尺寸,以及它在细胞中的大量数量,是线粒体转染的主要障碍。在这里,我们报道了利用线粒体靶向肽和细胞穿透肽的组合将外源DNA定位到拟南芥线粒体的细胞内递送。低浓度的肽足以将DNA传递到线粒体中,并且在短的孵育期(12小时)内,输入DNA的表达达到可检测的水平。我们发现,与细胞膜的静电相互作用并不是复杂内化的关键因素,相反,线粒体靶向复合物在细胞内渗透的改善显著提高了基因转移效率。我们的研究结果描述了一种简单有效的基于肽的方法,作为开发更复杂的植物线粒体转染策略的起点。
Available methods in plant genetic transformation are nuclear and plastid transformations because similar procedures have not yet been established for the mitochondria. The double membrane and small size of the organelle, in addition to its large population in cells, are major obstacles in mitochondrial transfection. Here we report the intracellular delivery of exogenous DNA localized to the mitochondria of Arabidopsis thaliana using a combination of mitochondria-targeting peptide and cell-penetrating peptide. Low concentrations of peptides were sufficient to deliver DNA into the mitochondria and expression of imported DNA reached detectable levels within a short incubation period (12 h). We found that electrostatic interaction with the cell membrane is not a critical factor for complex internalization, instead, improved intracellular penetration of mitochondria-targeted complexes significantly enhanced gene transfer efficiency. Our results delineate a simple and effective peptide-based method, as a starting point for the development of more sophisticated plant mitochondrial transfection strategies.
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