A PIGGYBAC TOOLKIT FOR RAPID TESTING OF PUTATIVE PEDIATRIC GBM DRIVER GENES WITH EXTENSIONS FOR INVESTIGATING CELLS OF RECURRENCE

We have created a novel method for studying glioma initiation, progression, and recurrence with in vivo-postnatal electroporation combined with piggyBac transposition. Electroporation allows for rapid addition of transgenes, simply by the inclusion of plasmid DNA in the initial injection. Thus, we have created a toolkit for interrogating the resulting tumor cells in vivo. Specifically, we have created piggyBac plasmids expressing a spectrum of transgenes or elements for modeling recently discovered and previously identified pediatric driver mutations, including H3.3, Atrx, Trp53, Nf1, and Pdgfra . Here we detail the use of these elements with additional transgenes for non-invasive imaging and inducible cell ablation. Further, we are investigating the use of novel methods for genetically labeling quiescent and recurrent cells, including inducible and reversible transgenic systems. Here we discuss these methodologies and their use in the context of our novel pediatric glioma models with a focus on emergent properties from different driver gene combinations.