Question

Describe an experiment designed to test whether the mouse gene Niere-1 is required for the normal development of the kidney during murine embryogenesis. You must explain the experimental controls you would use to interpret your results. How would you experimentally test whether the Niere-1 gene is sufficient to induce the formation of kidney in the mouse?

Answer 1

To test whether the mouse gene Niere-1 is required for the normal development of the kidney during murine embryogenesis there are following set of experiments are needed.

1) It is important to figure out the Niere-1 gene expression during the development. This can be done by using qPCR and western blot analysis of Niere-1 during kidney development

2) Inhibition of Niere-1 gene expression by using siRNA/RNA –interference/ embryo gene knockout strategy and studying its affect on the kidney development. Briefly, embryonic stem (ES) cells from early-stage mouse embryos can be used for gene targeting knock out experiment. This can be done by using the DNA that shares homologous sequence to the Niere-1 gene but has a reporter gene (For example GFP, green fluorescent protein) as insertion. This insertion will result in inactivated Niere-1 gene. Also, reporter gene can track the knock out mutation.

3) qPCR and western blot analysis of Niere-1 gene will confirm that Niere-1 gene is not functional in knockout embryo.