Analysis of a possible role of the mitochondrial Nipsnap family of proteins in the development of zebrafish.
Mitochondrial proteins play an immense role in the proper organogenesis of zebrafish. The Nipsnap family of proteins include three mitochondrial proteins (Nipsnap-1, -2 and -3), which have been shown to have a role in the neuronal organization of mice. The purpose of this study is to analyze a possible role of the Nipsnap proteins in the development of zebrafish by generating knock out strains for each of the three Nipsnap proteins using CRISPR/Cas9 technology.
We don’t know what type of distress the zebrafish lacking Nipsnap proteins would be in, but as NIPSNAP-1 knock-out mice are viable and fertile with no gross phenotype, we assume it would be mild to moderate distress.
The study would be very important in understanding the role of NIPSNAP proteins in vivo. It is of great importance to achieve an in depth understanding of disorders arising due to dysfunction of mitochondrial proteins – the molecular events happening and the disease mechanisms – in order to aim for better therapeutics, for example, Barth Syndrome, Pearson Syndrome, etc). We hope to use zebrafish as our animal model in order to answer some of our research questions on Nipsnap protein.
For a strong statistical significance and conclusion, we would have to work on at least 2000 zebrafishes under 7 days post fertilization (7dpf) for the whole project. To reduce the number of adults used, most of our experiments are performed on zebrafish larvae (before developmental stage requiring application), but we also need to perform some experiments on larvae under 7dpf. We have taken into consideration, the three R’s – replacement, reduction and refinement. Our initial studies have been on human cell lines. The animal experiments are important in concluding our in-vitro findings. We have reduced our estimated numbers considerably and 2000 7dpf zebrafish larvae are the bare minimum (for the whole project).
We don’t know what type of distress the zebrafish lacking Nipsnap proteins would be in, but as NIPSNAP-1 knock-out mice are viable and fertile with no gross phenotype, we assume it would be mild to moderate distress.
The study would be very important in understanding the role of NIPSNAP proteins in vivo. It is of great importance to achieve an in depth understanding of disorders arising due to dysfunction of mitochondrial proteins – the molecular events happening and the disease mechanisms – in order to aim for better therapeutics, for example, Barth Syndrome, Pearson Syndrome, etc). We hope to use zebrafish as our animal model in order to answer some of our research questions on Nipsnap protein.
For a strong statistical significance and conclusion, we would have to work on at least 2000 zebrafishes under 7 days post fertilization (7dpf) for the whole project. To reduce the number of adults used, most of our experiments are performed on zebrafish larvae (before developmental stage requiring application), but we also need to perform some experiments on larvae under 7dpf. We have taken into consideration, the three R’s – replacement, reduction and refinement. Our initial studies have been on human cell lines. The animal experiments are important in concluding our in-vitro findings. We have reduced our estimated numbers considerably and 2000 7dpf zebrafish larvae are the bare minimum (for the whole project).