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A gene delivery system for human cells mediated by both a cell-penetrating peptide and a piggyBac transposase

Paper ID Volume ID Publish Year Pages File Format Full-Text
7401 551 2011 13 PDF Available
Title
A gene delivery system for human cells mediated by both a cell-penetrating peptide and a piggyBac transposase
Abstract

The piggyBac (PB) transposable element has recently accumulated enormous attention as a tool for the transgenesis in various eukaryotic organisms. Arginine-rich cell-penetrating peptides (CPPs) are protein transduction domains containing a large amount of basic amino acids that were found to be capable of delivering biologically active macromolecules into living cells. In this study, we demonstrate a strategy, which we called “transposoduction”, which is a one-plasmid gene delivery system mediated by the nontoxic CPP-piggyBac transposase (CPP-PBase) fusion protein to accomplish both protein transduction and transposition. CPPs were proven to be able to synchronously deliver covalently linked PBase and noncovalently linked a cis plasmid into human cells. The expression of promoterless reporter genes coding for red (dTomato) and yellow (mOrange) fluorescent proteins (RFP and YFP) with PB elements could be detected in cells treated with the PBase-expressing plasmid after 3 days indicating transposition of coding regions to downstream of endogenous promoter sequences. An enhanced green fluorescent protein (EGFP) plasmid-based excision assay further confirmed the efficiency of the bifunctional CPP-PBase fusion protein. In conclusion, this strategy representing a combinational concept of both protein transduction and mobile transposition may provide tremendous potential for safe and efficient cell line transformation, gene therapy and functional genomics.

Keywords
Cell-penetrating peptide (CPP); Gene delivery; Polyarginine; piggyBac (PB); TransposonBGH, bovine growth hormone; CMV, cytomegalovirus; CNPT, covalent and noncovalent protein transductions; CPP, cell-penetrating peptide; EGFP, enhanced green fluorescent p
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A gene delivery system for human cells mediated by both a cell-penetrating peptide and a piggyBac transposase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 32, Issue 26, September 2011, Pages 6264–6276
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us