Block Copolymer/Plasmid DNA Micelles Postmodified with Functional Peptides via Thiol-Maleimide Conjugation for Efficient Gene Delivery into Plants
Block Copolymer/Plasmid DNA Micelles Postmodified with Functional Peptides via Thiol-Maleimide Conjugation for Efficient Gene Delivery into Plants
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DOI：
10.1021/acs.biomac.8b01304
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发表时间：
2019-02-01
影响因子：
6.2
通讯作者：
Numata, Keiji
中科院分区：
文献类型：
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作者：
Miyamoto, Takaaki;Tsuchiya, Kousuke;Numata, Keiji
将外源基因导入植物细胞对于农业和植物生物技术领域的广泛应用至关重要。具有细胞穿透和dna结合结构域的阳离子多肽载体成功地将外源基因传递到植物体内。然而，它们的细胞穿透活性可能会被细胞穿透肽（CPP）结构域和DNA货物之间不期望的静电相互作用减弱，从而导致基因传递效率有限。在这里，我们开发了马来酰亚胺共轭四（乙二醇）和聚（l -赖氨酸）（MAL-TEG-PLL）嵌段共聚物。通过与质粒DNA （pDNA）的静电相互作用，MAL-TEG-PLL形成胶束，在其表面呈现马来酰亚胺基团。胶束通过巯基-马来酰亚胺偶联实现了含半胱氨酸功能肽的后修饰，包括CPP （BP100-Cys）和核定位信号（Cys-NLS），从而避免了不必要的相互作用。通过对肽后修饰胶束的基因传递效率的比较，BP100-Cys在胶束表面的数量是基因传递效率的关键。与bp100预修饰胶束相比，bp100后修饰胶束的传递效率更高。因此，用功能性肽修饰聚合物胶束为设计高效的植物基因传递系统打开了大门。
Introducing exogenous genes into plant cells is essential for a wide range of applications in agriculture and plant biotechnology fields. Cationic peptide carriers with cell-penetrating and DNA-binding domains successfully deliver exogenous genes into plants. However, their cell-penetrating activity may be attenuated by undesired electrostatic interactions between the cell-penetrating peptide (CPP) domain and DNA cargo, resulting in limited gene delivery efficiency. Here, we developed the block copolymer maleimide-conjugated tetra(ethylene glycol) and poly(L-lysine) (MAL-TEG-PLL). Through electrostatic interactions with plasmid DNA (pDNA), MAL-TEG-PLL formed a micelle that presented maleimide groups on its surface. The micelle enabled postmodification with cysteine-containing functional peptides, including a CPP (BP100-Cys) and nuclear localization signal (Cys-NLS) via thiol-maleimide conjugation, thereby avoiding undesired interactions. According to a comparison of gene delivery efficiencies among the peptide-postmodified micelles, the amount of BP100-Cys on the micelle surface was key for efficient gene delivery. The BP100-postmodified micelle showed more efficient delivery compared with that of the BP100-premodified micelle. Thus, postmodification of polymeric micelles with functional peptides opens the door to designing highly efficient plant gene delivery systems.