Title

Dna Methylation Changes Resulting In Enhancer And Non-Enhancer Regions Of The Six3B Gene

Author(s)

Mackenzie Foster

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Mentor

Mentor: Ting Wang, Department of Genetics, Washington University in St. Louis

Abstract

Several regions upstream of genes affect whether transcription and translation occur. Two of these regions are the enhancer and the promoter, previously it was thought that promoters are the driving force for transcription, but now we are looking into the effects of enhancers. We isolated possible enhancer sequences, which we identified from where transcription factors bound, upstream of the six3b gene. These regions were cloned into entry and destination vectors for microinjection into zebrafish embryos. These regions where cloned so that expression of the GFP gene, which was located downstream of our possible enhancer regions, had the possibility to be expressed. If there was expression the region could be determined as an enhancer and if no expression was seen then the region could be looked at as a non-enhancer. After microinjection, we looked for patterned expression in the head and eyes of developing zebrafish, because the six3b gene has previously been shown to be expressed in the head and eyes region of the body. We identified the six3b-e1 region as a possible enhancer and the six3b-ne1 region as a possible non-enhancer, both by the fact that they were bound by transcription factors. The results provided evidence to support that these assumptions were correct because the six3b-e1 plasmid drove patterned expression in the head and eyes and the six3b-ne1 drove no patterned expression. We conclude that regions that experience DNA methylation, the addition of methyl groups to a cytosine, throughout development can be considered enhancer regions. Six3b-e1 experienced DNA methylation, but six3b-ne1 did not.

Start Date

4-11-2015 9:45 AM

End Date

4-11-2015 10:00 AM

COinS
 
Apr 11th, 9:45 AM Apr 11th, 10:00 AM

Dna Methylation Changes Resulting In Enhancer And Non-Enhancer Regions Of The Six3B Gene

Several regions upstream of genes affect whether transcription and translation occur. Two of these regions are the enhancer and the promoter, previously it was thought that promoters are the driving force for transcription, but now we are looking into the effects of enhancers. We isolated possible enhancer sequences, which we identified from where transcription factors bound, upstream of the six3b gene. These regions were cloned into entry and destination vectors for microinjection into zebrafish embryos. These regions where cloned so that expression of the GFP gene, which was located downstream of our possible enhancer regions, had the possibility to be expressed. If there was expression the region could be determined as an enhancer and if no expression was seen then the region could be looked at as a non-enhancer. After microinjection, we looked for patterned expression in the head and eyes of developing zebrafish, because the six3b gene has previously been shown to be expressed in the head and eyes region of the body. We identified the six3b-e1 region as a possible enhancer and the six3b-ne1 region as a possible non-enhancer, both by the fact that they were bound by transcription factors. The results provided evidence to support that these assumptions were correct because the six3b-e1 plasmid drove patterned expression in the head and eyes and the six3b-ne1 drove no patterned expression. We conclude that regions that experience DNA methylation, the addition of methyl groups to a cytosine, throughout development can be considered enhancer regions. Six3b-e1 experienced DNA methylation, but six3b-ne1 did not.