The beginning of this semester we continued our work on the C. elegans’ gene T20B12.7 and it was a little shaky due to the snowstorm and school closure. The first RNAi experiment we performed was unsuccessful because we could not get into the lab to record the results. We then performed another RNAi experiment, which show similar results as the previous semester. The RNAi T20B12.7 worms had only a small difference from the L440 worms in unhatched embryos. Another RNAi experiment was performed with rff-3 worms in the hope a more significant difference would be seen. The rff-3 worms are more sensitive to the RNAi treatment. However, the results from the RNAi experiments with rff-3 worms still had an insignificant difference. These results could have been for two different reasons. The first reason was that the RNAi was effecting the worms in another way besides embryo lethality. This was possible, but previous studies showed that this gene influences embryo development so not as likely as the second reason. The second reason for the insignificant difference could have been that something in our RNAi experiment was contaminated or defective. We decided since we were getting deterred by the RNAi experiments for T20B12.7, that we would hold on further experiments for this gene and complete experimentation on another gene, F55A3.3, that had had successful RNAi results.
F55A3.3 is a gene in C. elegans that is involved in embryo development, molting cycle, nucleus organization and reproduction. F55A3.3 is a human ortholog, spt16, that facilitates chromatin remodeling in the FACT complex. Previously performed RNAi experiments in our lab to knockdown F55A3.3 resulted in a sterile phenotype. We hypothesize that the loss of F55A3.3 may be necessary for normal embryo development. Our current experiments will analyze this hypothesis and help further understand the role this gene has in C. elegans. The next step was to use fluorescent microscopy to identify cause of sterility. We have performed two RNAi experiments and used microscopy to image the worms after they were treated. The first experiment we saw several abnormalities in the RNAi treated worms compared to the L44o worms. The abnormalities consisted of changed in the germline shape and irregular dividing in the embryos. The second RNAi experiment was not as successful. The imaging could not be performed properly. We are now in the process of troubleshooting the imaging process to see if the microscope is not working properly or if the worms that are tagged with GFP are contaminated.