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Cationic Cell Penetrating Peptide Modified SNARE Protein VAMP8 as Free Chains for Gene Delivery

Abstract

Previously, our group has carried out a series of work using branched polyethyleneimine with 25,000 g/mol molar mass (bPEI-25k) as gene delivery vectors. We came up with the theory that free cationic chains un-complexed with plasmid DNA (pDNA) can greatly promote gene transfection efficiency and influence the intracellular delivery process. These free chains can penetrate the membrane quickly, with some of them embedded inside the lipid bi-layers. The “stuck-out” cationic chain ends would shield the signal protein, prevent/delay the development of the later endolysosomes and enhance the efficiency of gene delivery. To mimic the effect of cationic polymers, we selected to use vesicle associated membrane protein-8 (VAMP8) and modified its N-terminus by different cationic cell penetrating peptides (CPPs). The modified fusion proteins are expressed in Escherichia Coli system and purified after extraction. These modified proteins are used as free chains for gene transfection with branched PEI to condense the pDNA. Results show that the gene transfection efficiency of PEI/pDNA polyplexes is obvious enhanced in 293T cell line, while the cell viability is fine. Further, the gene sequences encoding these modified VAMP8 were sub-cloned to pcDNA-3.1 and then transferred to 293T before PEI/pDNA polyplexes treatment. From the result, the transfection efficiency of PEI/pDNA complexes is improved by the similar level with that using modified proteins as free chains. Our current results further prove previous hypothesis that free cationic chains are probably embedded with membrane and influence intracellular trafficking, pointing out a new idea to design an effective non-viral gene deliver vector.

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Publication details

The article was received on 18 Jun 2018, accepted on 09 Aug 2018 and first published on 09 Aug 2018


Article type: Paper
DOI: 10.1039/C8BM00672E
Citation: Biomater. Sci., 2018, Accepted Manuscript
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    Cationic Cell Penetrating Peptide Modified SNARE Protein VAMP8 as Free Chains for Gene Delivery

    B. Chen and C. Wu, Biomater. Sci., 2018, Accepted Manuscript , DOI: 10.1039/C8BM00672E

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