Almost the end of the semester!
With the end of the semester approaching faster than anyone expected, the Williams lab team is both tying up loose ends and preparing for next steps in research.
Now that Alex has optimized the ChIP protocol, she will begin to either continue to research Ahr1 interactions or start our Nfe2 project. Quang has now identified certain genes from his bioinformatics project that he will be using next semester to do some molecular cloning with Larissa. Larissa will probes for in situ hybridization to further understand where these genes are localized, while Quang will inject capped mRNA for a rescue experiment; both project hope to further our understanding of how loss of nfe2 affects their expression. Jacob is still hard at work perfecting his microinjecting technique and has definitely seen improvement! He will be using this technique to knockdown Nrf3 with a morpholino to observe the developmental changes its loss induces.
Over the summer Larissa cross-bred two of the strains in lab, the nfe2 knockouts and the Brn3c’s (which have the GFP localized to their neuromasts). These were then raised up and this fall I have been genotyping the adult fish. Our hope was that we would get fish that were homozygous recessive knockouts (meaning both copies of nfe2 is mutant), but unfortunately most of the fish were either homozygous dominant or heterozygotes. This means that I will be joining Jacob in microinjecting Brn3c embryos second semester to observe the effects of Nfe2 loss on the development of neuromast cells. I also will be finishing up the SEM project with nfe2 knockout embryos before the semester ends.