Identification of Hox Regulatory Domains for Appendage Diversity and Fin-to-Limb Transition using Transgenic Zebrafish)
Session Number
C35
Advisor(s)
Tetsuya Nakamura, University of Chicago Neil Shubin, University of Chicago
Location
B-131 Grainger
Start Date
28-4-2016 11:05 AM
End Date
28-4-2016 11:30 AM
Abstract
To reveal the molecular mechanisms of appendage diversity and the fin-to-limb transition, I am identifying the regulatory domain of Hox expression in skate fin development. Hox genes are essential to organismal development but the genetic loci responsible for Hox expressions shifts remain elusive. Potential enhancer sites in the skate genome were identified using Assay for Transposase-Accessible Chromatin (ATAC) sequencing, cloned into the Green Fluorescent Protein (GFP) reporter vector and then injected into single-cell eggs of wild type zebrafish. To determine whether these enhancers were successfully integrated into the zebrafish genome, we used a polymerase chain reaction (PCR) to identify the presence of the GFP gene. In those that the PCR deemed GFP positive, second generation transgenic egg cells were examined for GFP fluorescence concentrated in the fin region. So far, I identified one transgenic fish; it showed GFP fluorescence in the hindbrain region. But I have yet to find transgenic fish showing fluorescence in the pectoral fin. I continue to identify transgenic fish by PCR and confirm localization of the GFP signal. Identifying these enhancers is not only indispensable in the study of ancient evolutionary mechanisms but also for the regulation of organ structure in regenerative biology and study of congenital diseases.
Identification of Hox Regulatory Domains for Appendage Diversity and Fin-to-Limb Transition using Transgenic Zebrafish)
B-131 Grainger
To reveal the molecular mechanisms of appendage diversity and the fin-to-limb transition, I am identifying the regulatory domain of Hox expression in skate fin development. Hox genes are essential to organismal development but the genetic loci responsible for Hox expressions shifts remain elusive. Potential enhancer sites in the skate genome were identified using Assay for Transposase-Accessible Chromatin (ATAC) sequencing, cloned into the Green Fluorescent Protein (GFP) reporter vector and then injected into single-cell eggs of wild type zebrafish. To determine whether these enhancers were successfully integrated into the zebrafish genome, we used a polymerase chain reaction (PCR) to identify the presence of the GFP gene. In those that the PCR deemed GFP positive, second generation transgenic egg cells were examined for GFP fluorescence concentrated in the fin region. So far, I identified one transgenic fish; it showed GFP fluorescence in the hindbrain region. But I have yet to find transgenic fish showing fluorescence in the pectoral fin. I continue to identify transgenic fish by PCR and confirm localization of the GFP signal. Identifying these enhancers is not only indispensable in the study of ancient evolutionary mechanisms but also for the regulation of organ structure in regenerative biology and study of congenital diseases.