Molecular Genetic Analysis of Congenital Stationary Night Blindness Associated TRPM1 Genetic Variants of Uncertain Significance in Humans and Horses Utilizing Caenorhabditis elegans
Date
2-14-2024
Faculty Mentor
Ashley Turner, Biology
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Submission Type
Conference Proceeding
Location
8:00-8:10am | Houston Cole Library, 11th Floor
Description
Congenital stationary night blindness (CSNB) is a collection of genetic diseases affecting the eyes and vision in humans and horses. In horses, genetic mutations in TRPM1 also result in a leopard-spotted coat pattern. Studies of Caenorhabditis elegans have revealed the nematode ortholog gon-2. If a TRPM1 missense variant has been associated with CSNB in humans and horses, then the genetic variant could result in gon-2 dysfunction and a phenotype in the mutant nematode. Identification of conserved variants of uncertain significance (VUS) within the TRPM1 were identified in both humans and horses. The human and horse TRPM1 genes, along with the nematode gon-2 gene were used to identify I875V VUS at the human TRPM1 location. Future experiments include generating a CRISPR-Cas9-engineered C. elegans model containing the TRPM1 missense VUS in the nematode loci of gon-2 for in vivo assessment. Primers were designed and used to amplify the I875V VUS region in C. elegans using polymerase chain reaction (PCR), gradient PCR, and gel electrophoresis. Gradient PCR was carried out to identify the optimal annealing temperature for a single band PCR product and a large PCR reaction was generated for an in vitro CRISPR-Cas9 experiment followed by microinjection. Ultimately, we plan to analyze phenotypic differences present in the mutant nematode strain when the gon-2 gene is edited with the TRPM1 VUS; focusing on the impact of gonadal and vulva development. This study will provide in vivo assessment of this CSNBassociated VUS shedding light on its clinical significance for humans and horses.
Keywords
student research, biology
Rights
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Disciplines
Biology
Recommended Citation
Moses, Victoria; Davis, Gabrielle; and Morris, Sara, "Molecular Genetic Analysis of Congenital Stationary Night Blindness Associated TRPM1 Genetic Variants of Uncertain Significance in Humans and Horses Utilizing Caenorhabditis elegans" (2024). JSU Student Symposium 2024. 39.
https://digitalcommons.jsu.edu/ce_jsustudentsymp_2024/39