Over the past year I have been developing and improving my research methods in order to get a better understanding of each steps involved in my project. So far we have moved from understanding that F10C2.4 is involved in reproduction to gathering data to support that F10C2.4 is not necessary in ovulation however is necessary to produce viable offspring. From this the results have shown that there is a dramatic decrease of progeny when F10C2.4 is not present. In preparing for presenting my research at the Society of Fellows I had to dig a lot more into my gene to see what it does. From that I have learned that it is a DNA Polymerase which may be necessary for specifically replication or something else. F10C2.4 is a gene that encodes for a protein which makes up to delta subunit of DNA Polymerase. From the schematic online, that looks like a small part of the overall Polymerase. It makes me think how such a small change can impact the viability of an organism.
I have learned so far that scientific research takes a lot of steps and preparation along with online research to understand what is updating within my field. However it has been a growing positive experience in which I hope to continue to develop my project.
Compared to last time, we have developed our methods of RNAi and developed a process which works for the way I want to express my experiment. Previously, I was having a hard time having consistent results but by this time around we have found a way to repeat the experiment and have consistent results. Along with this, I have learned how to make the bacteria to go on the plates, how to spot them and look at the worms under a microscope. With a trend of a large percentage of unhatched embryos with my gene of interest, we know that it must have some type of role in development. Under the microscope we looked at the adult mother worms from the RNAi plates, both the control (L440) and gene (F10C2.4) to look at physical differences. In order to do so, I used a different strain of worms. Instead of the wild type N2, I used a strain that had a GFP (green fluorescent protein) that showed the embryos and eggs in the mothers body which made it easier to see under the microscope. While looking at both types of worms, we realized that there was still something funky looking in the controls even though they showed normal progeny development. That made it difficult to compare what we saw to determine what is normal and what isn’t. This makes me realize that it is hard to have a “perfect” model to compare to which organisms are able to have mutations that may not effect the worm’s phenotype. As a result, I need to do more experiments and look at the worms under the microscope to see the physical differences. From that I can pin point the exact way I want to approach determining how this gene works.
The focus of my research is to determine the function of gene F10C2.4 in regards to Reproduction in C. elegans. C. elegans are microscopic worms that are model organisms since they have their whole genome sequenced, reproduces quickly, easy to maintain along with can easily be manipulated and observed due to its transparency. C. elegans have about 40% the same genes as humans, therefore we hope to compare our results to how reproduction occurs in humans. This project will help me practice the skills in learn in Genetics lab on a gene of my own thus furthering my understanding of the methods learned in class. The major method we have used is RNAi treatment which knocks out the gene of interest. So far, our results have not been consistent therefore we are tweaking factors in the worms and environment to see what gives us the best result. Overall, with the research papers I have read so far, we know very little about this gene. Hopefully, during this year I will be able to gather some more information about F10C2.4 in order to determine why we have this gene, how does it effect reproduction and whether or not my results transferable to human cells.