Several strategies have been developed to deliver exogenous macromolecules across cellular membranes. The intrinsic nature of cellular membranes precludes influx of macromolecules, such as DNAs, RNAs, and proteins. Small polar molecules, such as ions, amino acids, and monosaccharides, are able to enter cells through specific carriers or channels in the plasma membrane. Nitrogen (NH 3 +)/phosphate (PO 4 −) PBS, The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist.Įnhanced green fluorescent protein FITC,ġ-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan N/P, MOST 104-2320-B-259-002-MY3 (to H.-J.L.) from the Ministry of Science and Technology, Taiwan. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: Grant No. Received: DecemAccepted: FebruPublished: March 4, 2016Ĭopyright: © 2016 Liu et al. PLoS ONE 11(3):Įditor: Maxim Antopolsky, University of Helsinki, FINLAND This biomimetic LFcin L5a represents one of the shortest effective CPPs and could be a promising lead peptide with less immunogenic for DNA delivery in gene therapy.Ĭitation: Liu BR, Huang Y-W, Aronstam RS, Lee H-J (2016) Identification of a Short Cell-Penetrating Peptide from Bovine Lactoferricin for Intracellular Delivery of DNA in Human A549 Cells. The L5a and L5a/DNA complexes were not cytotoxic. Zeta-potential analysis revealed that positively electrostatic interactions of CPP/DNA complexes correlated with CPP-mediated transport. Treatment with calcium chloride increased the level of gene expression, without affecting CPP-mediated transfection efficiency. The L5a CPP delivered a plasmid containing the enhanced green fluorescent protein ( EGFP) coding sequence that was subsequently expressed in cells, as revealed by real-time PCR and fluorescent microscopy at the mRNA and protein levels, respectively.
Here we demonstrate that the L5a CPP (RRWQW) from bovine lactoferricin (LFcin), stably and noncovalently complexed with plasmid DNA and prepared at an optimal nitrogen/phosphate ratio of 12, is able to efficiently enter into human lung cancer A549 cells. Most of the CPP sequences in use today are based on non-native proteins that may be immunogenic. Cell-penetrating peptides (CPPs) have been shown to deliver cargos, including protein, DNA, RNA, and nanomaterials, in fully active forms into live cells.
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