Assessing the role of uncharacterised disease-causing candidate genes in zebrafish
In our laboratory we are trying to identify the genetic cause for developmental delay and intellectual disability identified in patients. To do so, we first screen genes previously associated with similar phenotypes however, in some cases, we are not able to find the disease-causing gene. In that case, we then analyse the patient’s entire genome and try to identify variants with potential to be causing the disease. From the list of candidate variants we start by choosing the most interesting candidates and assess their role in disease in zebrafish. To do so, zebrafish carrying mutations in these candidate genes are generated, usually using the CRISPR/Cas9 gene editing method, and then we analyse the phenotypes. In this research project we propose the analysis of zebrafish carrying mutations in 2 different genes associated with autism and ADHD. Stable mutant lines have already been generated but haven’t been characterised. We therefore give the opportunity for a student to carry on this research project, in a fairly independent fashion. This project gives the opportunity to the student to learn several techniques, from animal work to molecular biology techniques: handling zebrafish, crossing and possibly injecting embryos, genotype animals by PCR and Sanger sequencing, and phenotype analysis, by a combination of confocal microscopy, looking at the neuronal network, and behaviour analysis. |