Peptide-Based Polyion Complex Vesicles That Deliver Enzymes into Intact Plants To Provide Antibiotic Resistance without Genetic Modification
Peptide-Based Polyion Complex Vesicles That Deliver Enzymes into Intact Plants To Provide Antibiotic Resistance without Genetic Modification
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DOI：
10.1021/acs.biomac.0c01380
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发表时间：
2021-03-01
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：
Fujita，Seiya;Motoda，Yoko;Numata，Keiji
使用细胞穿透肽（CPP）将酶直接输送到完整植物中是一种无需基因改造即可改变植物功能的有吸引力的方法。然而，通过常规方法，由于酶在生理条件下会发生蛋白水解，因此难以长时间保持酶活性。在这里，我们开发了一种新型酶传递系统，使用聚离子复合囊泡（PICsomes）来保护酶免受蛋白酶的侵害。我们通过混合阴离子嵌段共聚物 alkyne-TEG-P(Lys-COOH) 和阳离子肽 P(Lys) 在表面创建了带有 PICsome 的反应基团。 PICsome 封装了新霉素磷酸转移酶 II (NPTII)（一种卡那霉素抗性酶），并在体外保护 NPTII 免受蛋白酶的影响。 CPP 修饰的 PICsome 将 NPTII 递送到拟南芥幼苗的根毛细胞中，并在幼苗中提供持续 7 天的卡那霉素抗性。因此，PICsome介导的酶传递系统是一种无需基因改造即可赋予植物长期瞬时性状的有前途的方法。
Direct delivery of enzymes into intact plants using cell-penetrating peptides (CPPs) is an attractive approach for modifying plant functions without genetic modification. However, by conventional methods, it is difficult to maintain the enzyme activity for a long time because of proteolysis of the enzymes under physiological conditions. Here, we developed a novel enzyme delivery system using polyion complex vesicles (PICsomes) to protect the enzyme from proteases. We created PICsome-bearing reactive groups at the surface by mixing an anionic block copolymer, alkyne-TEG-P(Lys-COOH), and a cationic peptide, P(Lys). The PICsome encapsulated neomycin phosphotransferase II (NPTII), a kanamycin resistance enzyme, and protected NPTII from proteases in vitro. A CPP-modified PICsome delivered NPTII into the root hair cells ofArabidopsis thalianaseedlings and provided kanamycin resistance in the seedlings that lasted for 7 days. Thus, the PICsome-mediated enzyme delivery system is a promising method for imparting long-term transient traits to plants without genetic modification.