During the summer 2019 and continuing into the school year, we worked in the developmental biology lab of Dr. Aaron Putzke here at Whitworth University. We worked on a project that was presented at an international conference by Dr. Putzke and that we presented at the regional Murdock College Science Research Conference in November of 2019. Our project has now been accepted to be presented by us at The Allied Genetics Conference in Washington DC in April. Our project has also led to a paper that we contributed to and is being submitted to the G3 journal of the Genetics Society of America.

            Dr. Putzke’s lab studies the function and role of a gene/protein called frk-1 (‘frk-1’refers to the gene, ‘FRK-1’ refers to the protein) in the species Caenorhabditis elegans. C. elegans is a small roundworm nematode that lives in soil and has been widely studied as a model of genetics. The frk-1 gene is very similar to the Fer kinase gene in humans and the Fer kinase gene has been shown to be important in multiple types of human cancers. This is the basis of our study of the frk-1 gene in C. elegans as we hope that our findings can be translated to humans. Dr. Putzke’s previous work has characterized the function of the FRK-1 protein in the cell as being important to regulating cell division during the early stages of development of the worm.

            A mutant worm strain was made where the frk-1 gene was removed from the DNA of the worms to see how it would affect the appearance and livelihood of the worms. This mutant strain of worms without the frk-1 gene showed many more defects than we expected given the prior knowledge of the FRK-1 protein’s function. Because of these we thought that the FRK-1 protein must have effects on other genes as parts of the worms were defective that the FRK-1 protein does not have a direct function in. In order to test this we analyzed the gene expression of all the genes in normal worms with frk-1 and in mutant worms without frk-1. Gene expression refers to the concept that different genes can be expressed and turned into proteins in some cells and not in others, and that amount of protein that is made can vary among cells as well. This gave us a way to see if the absence of frk-1 was affecting other genes. The lab performed the initial experiments (RNA isolation) before we started, but we helped with the later parts of these tests (quantitative PCR) for this step of the project. We found through our testing that over 2,000 genes were being expressed at different levels in the mutant worms compared to the normal worms. This showed that not only does FRK-1 have important roles in regulating cell division in early development, but it also has effects in several other parts of development though its regulation of the expression of many other genes.

            After we showed that FRK-1 affects the expression of many genes during development, we wanted to try to find other genes that might effect the expression of the frk-1 gene. In order to do this we performed RNA interference experiments on genes that we thought would be likely to affect frk-1 expression. RNA interference (RNAi) is a mechanism to prevent a gene from being expressed to make a protein without removing it from the DNA of the worms. Using this technique saved our lab a lot of time and money as compared to making a new mutant strain for every gene we tested. We are still working on screening each gene to look for effects in the expression of frk-1, but so far we have found several genes that have indeed affected frk-1 expression. In order to visualize the FRK-1 protein during these RNAi experiments we use microscopy with a strain of worms that glows wherever FRK-1 is expressed.