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Updated: May 2, 2026

piggyBac Transposon System Modification of Primary Human T Cells

piggyBac Transposon System Modification of Primary Human T Cells

Published on: November 5, 2012

An optimized polymeric delivery system for piggyBac transposition.

Daniel Nisakar Meenakshi Sundaram¹, Remant Bahadur K C¹, Wei Fu²

¹Department of Chemical and Materials Engineering, Faculty of Engineering, University of Alberta, Edmonton, Alberta, Canada.

Biotechnology and Bioengineering

|February 19, 2024

Summary

A novel polymeric gene delivery system using piggyBac transposition enables stable, long-term gene expression. This method, utilizing polyethylenimine polymer with lipid substitution (PEI-L), offers a promising alternative to viral gene delivery for therapeutic applications.

Keywords:

mRNA piggyBac polyethylenimine stable gene expression transposase transposon

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Published on: January 12, 2018

Area of Science:

Gene therapy
Molecular biology
Biotechnology

Background:

The piggyBac transposon/transposase system is a promising tool for stable gene expression via genomic integration.
Current delivery methods for piggyBac components primarily rely on physical transfection, with limited exploration of polymeric systems.
Viral vectors, while effective, present safety concerns and limitations for long-term gene therapy.

Purpose of the Study:

To develop and optimize an effective polymeric gene delivery system for the piggyBac transposon/transposase system.
To achieve long-term, stable transgene expression using a non-viral delivery method.
To evaluate the efficiency and stability of piggyBac-mediated gene delivery using a novel polyethylenimine polymer with lipid substitution (PEI-L).

Main Methods:

Development of a single-nanocomplex using low molecular weight polyethylenimine polymer with lipid substitution (PEI-L).
Formulation encapsulated three components: piggyBac transposon plasmid DNA (PB-GFP), piggyBac transposase (plasmid or mRNA), and polyacrylic acid additive.
Transfection and assessment of stable green fluorescence protein (GFP) expression in MDA-MB-231 and SUM149 cell lines.

Main Results:

Optimized PEI-L formulation achieved stable GFP expression in MDA-MB-231 cells for up to 108 days and in SUM149 cells for up to 43 days after a single treatment.
High transgene stability was observed, with expression retained after three consecutive cryopreservation cycles.
Efficient delivery of all three piggyBac components was achieved in a single complex, using a low starting cell number.

Conclusions:

A novel polymeric delivery system (PEI-L) effectively facilitates piggyBac-mediated stable gene expression.
This non-viral approach offers a viable and potentially safer alternative to viral transduction for gene therapy applications.
The demonstrated stability and efficiency highlight the potential of PEI-L for future therapeutic gene delivery strategies.