Development of Novel Tools for Radial Glia Lineage Tracing and Modeling Central Nervous System Tumors

In this dissertation, I first established a novel non viral piggyBac transposon mediated transgenesis approach delivered by in utero electroporation for radial glia lineage tracing. Furthermore, I developed a novel central nervous system tumor model in rat using piggyBac IUE. By directing expression of oncogenic HRasV12/AKT in radial glia, astrocytes and oligodendrocytes, glioblastoma multiforme and anaplastic oligoastrocytoma were induced. Also addition of neurogenic bHLH transcription factors Neurogenin 2 and Neural differentiation 1 to GBM inducing HRasV12/AKT, malignant pediatric atyical teratoid rhabdoid tumor like tumor was induced. These results shad lights on the sources of tumor heterogeneity and indicate that oncogenic event occurring in disparate cell types and/or molecular context can lead to different tumor formation. Last, I explored the application of RNA guided genome engineering tool CRISPR/Cas9 in studying necortical development and model human neural developmental disorder as well as glioblastoma multiforme using CRISPR/Cas9 system. In utero electroporation of CRISPR/Cas9 constructs targeting PTEN, successfully knocked out PTEN expression in neurons. PTEN negative neurons showed hypertrophy and decreased resting membrane potential, increased sEPSCs and mEPSCs. Combinations of gRNAs targeting PTEN, NF1 and P53 induced glioblastoma multiforme. These new tools shall be proven to be powerful to study neocortical development and central nervous system tumors.