4.7 Article

Peptide nanostructures-based delivery of DNA nanomaterial therapeutics for regulating gene expression

期刊

MOLECULAR THERAPY-NUCLEIC ACIDS
卷 33, 期 -, 页码 493-510

出版社

CELL PRESS
DOI: 10.1016/j.omtn.2023.07.017

关键词

DNA nanostructures; DNA nanotechnology; MT: Oligonucleotides: Therapies and Applications; VEGF; delivery; nucleic acid therapeutics; peptide; self-assembly

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This study demonstrates the use of self-assembled linear peptides as delivery agents for functional bDNA nanomaterials in mammalian cells. The W4R4 peptides show excellent internalization of antisense bDNAs and significantly suppress the expression of VEGF.
Self-assembled branched DNA (bDNA) nanomaterials have exhibited their functionality in various biomedical and diagnostic applications. However, the anionic cellular membrane has restricted the movement of bDNA nanostructures. Recently, amphiphilic peptides have been investigated as cationic delivery agents for nucleic acids. Herein, we demonstrate a strategy for delivering functional bDNA nanomaterials into mammalian cells using self-assembled linear peptides. In this study, antisense oligonucleotides of vascular endothelial growth factor (VEGF) were inserted in the overhangs of bDNAs. Novel linear peptides have been synthesized and the peptide-bound bDNA complex formation was examined using various biophysical experiments. Interestingly, the W4R4-bound bDNAs were found to be exceptionally stable against DNase I compared to other complexes. The delivery of fluorescent-labeled bDNAs into the mammalian cells confirmed the potential of peptide transporters. Furthermore, the functional efficacy of the peptide-bound bDNAs has been examined through RT-PCR and western blot analysis. The observed results revealed that W4R4 peptides exhibited excellent internalization of antisense bDNAs and significantly suppressed (3- to 4-fold) the transcripts and translated product of VEGF compared to the control. In summary, the results highlight the potential use of peptide-based nanocarrier for delivering bDNA nanostructures to regulate the gene expression in cell lines.

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