Epilepsy is a devastating neurological disorder characterized by incontrollable synchronous neuronal activity, which clinically manifests as epileptic seizures. Epilepsy usually develops after an initial insult, such as head trauma or stroke, which triggers the epileptogenic process. During epileptogenesis, the affected brain regions undergo multiple changes in connectivity and gene expression, and between the most affected genes are those responsible for network inhibition. These include genes coding for GABA receptors, which are usually downregulated during epileptogenesis. Recent developments in gene editing technologies, including the bacteria-derived CRISPR-Cas9 system, have opened new avenues for gene therapy. In particular, the possibility of developing a reversible and precise regulatory system for endogenous genes with high spatio-temporal control, overcomes several limitations of the classic gene therapy approaches. This project aims to apply an on-demand, controllable gene expression technology, based on guide RNAs and a catalytically inactive Cas9, targeting the expression of GABA receptors for the control of epileptogenesis. We will first employ a system for inducible control of GABA receptor expression in neuronal cultures, verify its efficiency with electrophysiology, molecular biology, immunohistochemistry and super-resolution microscopy, and finally apply it to animal models of epilepsy, with the aim of blocking the development of chronic epilepsy.
|Effective start/end date||2016/01/01 → 2019/12/31|