Generation and characterization of new mouse models and transcription profiling

• Applicant: Privatdozent Dr. Thomas Ott
• Subject Area: Molecular and Cellular Neurology and Neuropathology ; Molecular Biology and Physiology of Neurons and Glial Cells
• Term: since 2017
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 377782854

Due to the complexity of the genetic epileptic disorders and the lack of knowledge of the underlying mechanisms in brain networks, deeper analyses are needed to evaluate the functional consequences of mutations for the regulation of gene expression and signaling cascades. A major focus of this project will be the generation and phenotypic characterization of mouse knock-in (KI) models designed to characterize known epileptogenic mutations in vivo. Initially, KCNA2 (P6) and SCN8A (P5), two examples of newly identified epilepsy-associated genes encoding for voltage-gated ion channels, which (i) are expressed in both excitatory and inhibitory neurons and (ii) carry gain-of-function as well as loss-of-function mutations causing epilepsy and intellectual disability, will be selected to generate conventional KI mutant lines expressing the missense mutation and conditional KI transgenic mouse lines, allowing us to achieve a recombinase-dependent time- or region-specific expression of the mutant alleles. Additional models will be generated for two to three newly identified variants from projects P1-3 that are most interesting with respect to the disease phenotype and affected gene. Another major focus of this project is to establish region- and cell population-specific gene expression profiles in mouse models of genetic epilepsies to obtain a more detailed picture of transcriptional changes during epileptogenesis, which will be correlated with human genetic findings in projects P1-P3 and physiological changes observed in P4-P8. To this end, we will take advantage of latest developments in single-cell transcriptomics and apply novel methods such as microfluidic droplet generation or dispension into nano-well chips to isolate hundreds to thousands of single brain cells for highest resolution regarding the contribution of particular cell types to epileptic seizures.

• DFG Programme: Research Units
• Subproject of: FOR 2715: Epileptogenesis of genetic epilepsies