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Intracellular Delivery of DNA and Protein by a Novel Cell-Permeable Peptide Derived from DOT1L.

Authors
  • Geng, Jingping1, 2
  • Guo, Xiangli1, 2
  • Wang, Lidan1, 2
  • Nguyen, Richard Q3
  • Wang, Fengqin2
  • Liu, Changbai2
  • Wang, Hu1, 3
  • 1 Department of Pathology and Immunology, Medical School, China Three Gorges University, Yichang 443002, China. , (China)
  • 2 Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, China Three Gorges University, Yichang 443002, China. , (China)
  • 3 Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Type
Published Article
Journal
Biomolecules
Publisher
MDPI AG
Publication Date
Feb 02, 2020
Volume
10
Issue
2
Identifiers
DOI: 10.3390/biom10020217
PMID: 32024261
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Cellular uptake and intracellular release efficiency of biomacromolecules is low because of hurdles in the cell membrane that result in limited access to intra-cellular targets with few functional effects. Cell-penetrating peptides (CPPs) act as cargo delivery vehicles to promote therapeutic molecule translocation. Here, we describe the novel CPP-Dot1l that not only penetrates by itself, but also mediates cargo translocation in cultured cells, as confirmed by fluorescence microscopy and fluorescence spectrophotometry. We conducted cytotoxicity assays and safety evaluations, and determined peptide-membrane interactions to understand the possible pathway for cargo translocation. Additional nucleic acid and covalently conjugated green fluorescence protein (GFP) studies mediated by CPP-Dot1l were conducted to show functional delivery potential. Results indicate that CPP-Dot1l is a novel and effective CPP due to its good penetrating properties in different cell lines and its ability to enter cells in a concentration-dependent manner. Its penetration efficiency can be prompted by DMSO pretreatment. In addition, not only can it mediate plasmid delivery, but CPP-Dot1l can also deliver GFP protein into cytosol. In conclusion, the findings of this study showed CPP-Dot1l is an attractive pharmaceutical and biochemical tool for future drug, regenerative medicine, cell therapy, gene therapy, and gene editing-based therapy development.

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