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A Synthetic DNA Delivery System Mediated by Arginine-Rich Cell-Penetrating Peptides in the Kingdoms Animalia, Plantae and Protista pp. 363-372 $100.00
Authors:  (Yun-Hao Dai, Yung-Jen Chen, Min-Jie Liu, Han-Jung Lee, Department of Natural Resources and Environmental Studies, National Dong Hwa University, Hualien, Taiwan)
Abstract:
Delivery of synthetic DNA into cells is crucial in biotechnology. Nonviral DNA delivery systems have become increasingly desirable in DNA transfer nowadays due to ease of assembly and low levels of toxicity, immunogenicity and insertional mutagenesis. Cell-penetrating peptides (CPPs), also called protein transduction domains (PTDs) or arginine-rich intracellular delivery (AID) peptides in our group, were small peptides rich in basic amino acids and were proven to be required for the translocation through the plasma membrane and the entry into cells. In recent years, we demonstrated that arginine-rich CPPs are able to not only penetrate cell membranes but also carry cargoes, such as proteins, RNAs, DNAs and nanoparticles, into living cells in different manners. However, our study of DNA delivery mediated by CPPs was limited in plant cells initially. Herein, we selected various cells of representative species from the kingdoms animalia, plantae and protista to test whether CPPs are able to carry plasmid DNAs into cells. CPPs were applied to deliver the fluorescence-labeled DNA and the fluorescent reporter gene into different cells by a noncovalent manner. The shuttle of image and protein products of gene expression were detected by a fluorescent microscope. We found CPPs could deliver DNAs into cells of insect, mung bean and paramecia. Finally, we confirmed that CPPs showed no cytotoxicity by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diohenyltetrazolium bromide (MTT) assay. In the future, this synthetic CPP-mediated DNA delivery system will be an efficient and safe tool of gene transfer in basic bioscience research and clinical therapies. 


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A Synthetic DNA Delivery System Mediated by Arginine-Rich Cell-Penetrating Peptides in the Kingdoms Animalia, Plantae and Protista pp. 363-372