Research AREA: Targeted CRISPR-Cas Delivery into Mammalian Nervous System
Research Description
The discovery of the CRISPR-Cas system and its application to specifically edit the genes in vivo provides new opportunities to treat neuropathologies at the genome level. A set of essential aspects need to be addressed to establish an effective approach to CRISPR-based gene editing in the mammalian brain. Most effective CRISPR-Cas systems have to be screened, appropriate vectors for CRISPR-Cas delivery have to be selected, their efficiency has to be trialled in the living brain, and the outcome of selective gene editing needs to be thoroughly assessed. We approach this challenging task in a stepwise manner to develop a CRISPR-Cas delivery system for selective gene editing of brain cells and to apply it in the models of nervous system disorders.
We use these new tools to define the molecular signalling pathways that drive this highly specific pruning of unnecessary synapses. We use both ex vivo tissue cultures and genetically modified mouse lines for this. We are developing novel molecular tools for rapid, selective and sensitive labelling of synaptic surface molecules. High-resolution fluorescent microscopy of developing circuits is supplemented with electrophysiology and animal behaviour experiments. We intend to define the synapses destined for elimination in vitro and thereafter in vivo and
elucidate their molecular signatures, giving first direct insights into the molecular cascades
required for developmental synaptic pruning in the maturing circuits of the brain.
