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| Main Authors: | , , , |
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| Format: | Artículo Open Access |
| Published: |
Wiley
2024
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| Subjects: | |
| Online Access: | https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bit.28665 |
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Table of Contents:
- An optimized polymeric delivery system for piggyBac transposition Daniel Nisakar Meenakshi Sundaram Remant Bahadur K. C. Wei Fu Hasan Uludağ Biotechnology and Bioengineering AbstractThe piggyBac transposon/transposase system has been explored for long‐term, stable gene expression to execute genomic integration of therapeutic genes, thus emerging as a strong alternative to viral transduction. Most studies with piggyBac transposition have employed physical methods for successful delivery of the necessary components of the piggyBac system into the cells. Very few studies have explored polymeric gene delivery systems. In this short communication, we report an effective delivery system based on low molecular polyethylenimine polymer with lipid substitution (PEI‐L) capable of delivering three components, (i) a piggyBac transposon plasmid DNA carrying a gene encoding green fluorescence protein (PB‐GFP), (ii) a piggyBac transposase plasmid DNA or mRNA, and (iii) a 2 kDa polyacrylic acid as additive for transfection enhancement, all in a single complex. We demonstrate an optimized formulation for stable GFP expression in two model cell lines, MDA‐MB‐231 and SUM149 recorded till day 108 (3.5 months) and day 43 (1.4 months), respectively, following a single treatment with very low cell number as starting material. Moreover, the stability of the transgene (GFP) expression mediated by piggyBac/PEI‐L transposition was retained following three consecutive cryopreservation cycles. The success of this study highlights the feasibility and potential of employing a polymeric delivery system to obtain piggyBac‐based stable expression of therapeutic genes. 10.1002/bit.28665 http://onlinelibrary.wiley.com/termsAndConditions#vor