Hoballah, Talal; Martinez , Angela
College of Science and Mathematics
Professor: Dr. Rachel Hopp
ABSTRACT:
Cep-1 knockdown in C. elegans using RNAi and the effects of DNA damaged induced apoptosis
As a cell progresses through the cell cycle, the cell goes through various check points. These check points are necessary to prevent mutations in their DNA due to stresses such as UV radiation. When a cell with DNA damages is detected by the G2/M check point it is removed by various methods. One of these methods is through apoptosis, which is programmed cell death that successfully removes the DNA damaged cell. DNA damaged cells may become cancerous cells which may explain why they have unregulated cell division. One way this problem is combated is through p53, a tumor suppressing gene in humans, which when activated can lead to apoptosis. This process has been studied in model organisms such as Caenorhabditis elegans. C. elegans have a similar gene called cep-1 and when activated can lead to apoptosis when cells with DNA damaged have been detected. To further examine this phenomenon, it was decided to explore this process of apoptosis in C. elegans using RNAi. RNAi is a gene suppressing mechanism that can inhibit the expression of specific proteins used in gene expression. In this experiment, wild type N2 C. elegans worms were fed IPTG induced E. coli HT115 (DE3)/ PPR244 (F52B5.5), which has the RNAi target for cep-1. The control group was fed with the same bacteria, but RNAi was not induced with IPTG. Both groups of N2 worms are exposed to a 5 minutes to 2 hours range of UV light that has the wavelength of between 300 and 400 nm. After irradiation, the C. elegans with cep-1 knockdown were transferred to new NGM + kanamycin plates every 24 hours during egg laying period. After transferring, N2 C. elegans are examined under a dissecting microscope and were manually counted and the hatching percentage is calculated. By conducting these types of experiments, scientist can further under p53 role in combating cancer in humans. Results will be presented at symposium.
