Event Title

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

Disciplines

Biology

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.


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Apr 28th, 11:05 AM Apr 28th, 11:30 AM

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.