What causes PCDH19-epilepsy?
This honours projects seeks to develop a stronger understanding of the pathological mechanism of PCDH19-girls clustering epilepsy.
Mutations in the X-linked PCDH19 gene are a leading cause of genetic epilepsy. PCDH19-Girls Clustering Epilepsy (PCDH19- GCE) has a unique form of X-linked inheritance as carrier (heterozygous) females are affected while carrier (hemizygous) males are not.
Affected girls suffer from seizures of variable severity and some also have intellectual disability and/or autism. PCDH19 is a transmembrane protein that enables cells to ‘stick together’ during brain development.
Using CRISPR mouse models, we have recently shown that PCDH19 heterozygous female mice have elevated brain activity and that the neurons cluster abnormally during development.
The aim of this project is understand how PCDH19 mutations cause epilepsy at the cellular and molecular level. We will firstly use fluorescence immunodetection methods to identify the neural subtypes that express PCDH19.
Using conditional mouse models and cutting-edge in utero electroporation, we will delete PCDH19 from specific neuronal cells and assess the impact on brain activity and development. Primary neuronal cultures of normal and mutant cells will also be used to determine PCDH19 function in synapse formation and neuronal activity.
Research techniques include mouse handling, PCR, cell culture, histology and immunofluorescence.
Study genome editing
CRISPR genome editing technology is transforming medicine, biology and agriculture. CRISPR enables targeted genetic modification of virtually any species with unprecedented efficiency. Given its potential, CRISPR is envisioned to be a game changer for therapeutic development, particularly for incurable genetic diseases.
The Genome Editing Laboratory led by Professor Paul Thomas uses state-of-the-art molecular genetic approaches to develop CRISPR technology to enhance human health. Our CRISPR innovation includes the development of strategies to eliminate entire chromosomes that could potentially be deployed for treatment of aneuploidy diseases.
Our lab is expert in generating genetically modified mice using CRISPR to model human disease mutations with >60 mouse models to date. We are using these unique models to investigate the pathology of relatively common genetic diseases such as epilepsy and muscular dystrophy. We are also developing CRISPR genome editing approaches to cure genetic diseases by correcting disease-causing mutations in vivo.
Finally, we are leading the world in the development of CRISPR gene drives in mice. This powerful technology has enormous potential for controlling invasive mouse populations that spread (zoonotic) disease, cause species extinction and loss of agricultural productivity.