PiggyBac transposon-based gene delivery with cationic ultrasound contrast agents into SCID and CD-1 mice.

Abstract

Efficient integration of a functional gene, along with suitable transcriptional regulatory elements, poses a fundamental difficulty for the application of successful gene therapy. Though inducible gene expression after somatic cell gene transfer has been achieved by employing viral vectors, issues associated with cargo capacity, host immune response, the risk of insertional mutagenesis, and the requirement for separate viruses are limiting factors. Several researchers have used transposon-based approaches for gene delivery, and the piggyBac transposon system has recently been shown to be effective in human cell lines and for the transgenesis in mice. The use of engineering helper-independent plasmids with the mouse codon-biased piggyBac transposon (mPB) gene in combination with cationic ultrasound contrast agents (UCAs) is investigated for a range of insonification parameters. The plasmid pmGENIE-3 deactivates the mPB gene after insertion of the transposon, eliminating potentially negative effects which may occur from the persistence of an active piggyBac gene post-transposition.