Cell-penetrating peptide-mediated transfection of proteins into tissues of Arabidopsis and Chinese cabbage

文献类型： 外文期刊

第一作者： Wu, Han

作者： Wu, Han;Zhao, Ying;Zhang, Yun;Feng, Hui;Wu, Han;Zhang, Zhipeng;Zhu, Kai;Wang, Yanqiu;Ke, Fulai;Zhang, Fei

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关键词： Cell-penetrating peptides; Genome editing; DNA-free; Chinese cabbage; Arabidopsis

期刊名称：IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT （ 影响因子：2.347； 五年影响因子：2.404 ）

ISSN： 1054-5476

年卷期： 2022 年 58 卷 1 期

页码：

收录情况： SCI

摘要： Cell-penetrating peptides (CPPs) are a class of short peptides, which have the ability of intracellular delivery. CPPs can also be used as an ideal transfection tool whereby biomacromolecules can be transferred as cargo into recipient cells. Here, a synthesized cell-penetrating peptide (R9)-mediated DNA-free transfection is presented. DNA sequences of nine tandem arginine (R9), one cysteine (cys), reporter mCherry, and histidine label were sequentially constructed into pET-45b( +) expression vector and transformed into the Escherichia coli BL21(DE3) strain. The R9-cys-mCherry fusion protein was obtained by prokaryotic expression. The Arabidopsis root tips, Chinese cabbage microspores, and 3-wk-old microspore embryos (MDEs) were used as transfected recipients. With a working concentration of R9-cys-mCherry ranging from 10 to 100 mu g mL(-1) and overnight incubation, R9-cys-mCherry protein can be translocated into the recipient mentioned above. The transfection efficiency of root tips reached 100%, and that of microspores and MDEs was 8.13% and 94.79%, respectively. A CPP-mediated DNA-free transfection system was built in dicots providing a technical basis for DNA-free CRISPR RNP (ribonucleoprotein) intracellular delivery in the future.

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