Characterization of CRISPR/CAS9-Mediated Cabin1 Deletion on Craniofacial Morphology in Larval Zebrafish
Mentor 1
Dr. Ava Udvadia
Location
Union Wisconsin Room
Start Date
28-4-2017 1:30 PM
End Date
28-4-2017 4:00 PM
Description
Although craniofacial abnormalities are among the most common birth defects, the genetic mechanisms regulating craniofacial development remain poorly understood. We are interested in exploring novel roles of a transcriptional regulator originally discovered for its role in T-cell activation, Cabin1, in craniofacial development. Based on our preliminary findings we hypothesize that Cabin1 is necessary for appropriate craniofacial development, including proper migration and differentiation of the cranial neural crest cells. To test our hypothesis, we collaborated with Dr. Brian Link (Medical College of Wisconsin) to generate Cabin1 knockout fish using CRISPR/Cas9 genome editing. We have two independent mutant strains in which the Cabin1 gene contains a premature stop codon and is expected to give rise to functionally null proteins. Here we will present our preliminary findings on the morphological differences in craniofacial development between Cabin1 mutant and wildtype strains. To characterize the developing craniofacial cartilage, we stained 5-day old larvae with Alcian Blue and used morphometric analysis to identify differences in craniofacial morphology between Cabin1 mutants and wildtype strains. Our preliminary analyses suggest that there are differences in jaw morphology of the Cabin1 mutants, however the differences are subtle compared with those we previously observed using gene knockdown strategies. We are currently investigating how the observed craniofacial phenotypes in our mutant strains correlate with long term survival.
Characterization of CRISPR/CAS9-Mediated Cabin1 Deletion on Craniofacial Morphology in Larval Zebrafish
Union Wisconsin Room
Although craniofacial abnormalities are among the most common birth defects, the genetic mechanisms regulating craniofacial development remain poorly understood. We are interested in exploring novel roles of a transcriptional regulator originally discovered for its role in T-cell activation, Cabin1, in craniofacial development. Based on our preliminary findings we hypothesize that Cabin1 is necessary for appropriate craniofacial development, including proper migration and differentiation of the cranial neural crest cells. To test our hypothesis, we collaborated with Dr. Brian Link (Medical College of Wisconsin) to generate Cabin1 knockout fish using CRISPR/Cas9 genome editing. We have two independent mutant strains in which the Cabin1 gene contains a premature stop codon and is expected to give rise to functionally null proteins. Here we will present our preliminary findings on the morphological differences in craniofacial development between Cabin1 mutant and wildtype strains. To characterize the developing craniofacial cartilage, we stained 5-day old larvae with Alcian Blue and used morphometric analysis to identify differences in craniofacial morphology between Cabin1 mutants and wildtype strains. Our preliminary analyses suggest that there are differences in jaw morphology of the Cabin1 mutants, however the differences are subtle compared with those we previously observed using gene knockdown strategies. We are currently investigating how the observed craniofacial phenotypes in our mutant strains correlate with long term survival.
