Research Description

E2F6 mediated DNA Methylation

I research gene regulation mediated by E2F6, a transcription factor which regulated meiosis specific genes. I am specifically trying to determine if E2F6 is regulating these genes through DNA methylation. DNA methylation regulates genetic expression because when a methyl group attaches to a cytosine that is adjacent to a guanine (a CpG site) the DNA hypercoils and cannot be transcribed. If a gene cannot be transcribed then the protein will never be formed and the gene is effectively turned off. Gene regulation is an important area of health research because the cause of many diseases is faulty regulation. Faulty regulation of meiotic genes in somatic tissue (like the ones in this study) can cause aneuploidy. Aneuploidy is the term for a cell with the incorrect number of chromosomes, and is often the cause of cancer. This research will offer new understanding in carcinogenesis.

I am examining the role of E2F6 in DNA methylation by comparing methylation patterns of meiosis specific genes in wildtype and E2F6 deleted MEFs. In order to achieve this I must first extract DNA from both wildtype MEFs and E2F6-null MEFs. I then bisulfite treat the DNA so that any unmethylated cytosines are converted to thymines. Once the DNA is converted I amplify the promoter region (including the E2F6 binding site) of specific genes by PCR. I then clone this DNA into plasmids and sequence the results. Once I have sequenced the genes I can compare the results to the original gene sequence. This comparison shows how many CpG sites have been methylated. I can then compare the E2F6 wildtype MEFs to the E2F6-null MEFs, which provides further understanding into the role of E2F6 on DNA methylation.