Nisa Habibi
Pronouns:
Research Mentor(s): Yuji Mishina
Research Mentor School/College/Department: Biological and Material Sciences / Dentistry
Program:
Authors: Nisa Habibi, Yuji Mishina
Session: Session 4: 1:40 pm – 2:30 pm
Poster: 102
Abstract
Ellis-van Creveld (EvC) syndrome is an autosomal recessive disease characterized with chondrodysplasia dwarfism, a form of dwarfism which results in abnormal bone development, causing the shortened development of limbs, in addition to abnormal oral cavity development. This disease is rare with an occurrence of about 1 in 60,000 (Zhang et al. 2015). The syndrome’s phenotypic expression includes shortening of limbs, short ribs, and difficulty breathing. The EVC2 gene, situated many mutations including the ones within exon 12, has been identified as a causative gene for this syndrome, yet its precise role in growth factor signaling including that of Hedgehog —a pathway vital for regulating embryonic development—remains ill-defined. Given the limited understanding of how EVC2 mutations contribute to EvC syndrome, it is vital to develop a mouse line to explore how changes in the Evc2 gene can lead to EvC syndrome. Previously, Mishina Lab worked to generate conventional and conditional mutant Evc2/Limbin alleles. These mutant mice show interesting cartilage defects including the knee joint, the skull base synchondrosis, the condyle head in the temporomandibular joint, and the tracheal cartilage, which I am focusing as my UROP project. To distinguish mutant mice from controls, forward and reverse primers, LbnFR1 and LbnRev3 respectively, were designed and have been used to target a region of exon 12 within the genomic sequence, to identify the presence of the wildtype allele for this gene. The mutant allele was genetically engineered, and a separate reverse primer called LbnC was designed to specifically identify the mutant allele, generating a shorter PCR product. Thus, LbnFR1 and LbnRev3 serve to target the wildtype sequence for the expected allele, whereas LbnFR1 and LbnC serve to target the sequence of this proposed mutant allele. Together, these primers compared the wildtype from the mutant, where the mouse line undergoes the premature termination of EVC2 protein, mimicking mutations found in EvC patients (Zhang et al. 2015). However, recent challenges with PCR assays necessitate the development of new primers for a more effective modeling of the Evc2/Limbin allele. This project seeks to generate these new primers using Primer Blast software, with the efficacy of these primers to be evaluated through PCR and gel electrophoresis analysis. By refining the method of detecting the genetic modeling of Evc2 mutations, this research aims not only to deepen our understanding of EvC syndrome’s pathogenesis but also to lay the groundwork for potential therapeutic interventions.”
