Finn Topham1,2, Shohei Takaoka2, Xiangbo Ruan2

Eckerd College1, Institute for Fundamental Biomedical Research Johns Hopkins All Children's Hospital2

ZNF408 gene mutations are linked to retinal degeneration, however little is known about how these ZNF408 mutations cause diseases. ZNF408 mutations S126N and H455Y and other missense variants of interest linked with Retinitis Pigmentosa were identified in an NCBI search. PCR site-directed mutagenesis was performed to clone the mutants into mammalian expression plasmids. To investigate the downstream effects of the selected mutations, plasmids carrying wild-type ZNF408 or ZNF408 mutants were transfected into human retinal pigment epithelial ARPE-19 cells. Silver staining and immunoprecipitation experiments were performed to determine how ZNF408 mutants affect the interaction between cellular proteins and ZNF408. Immunofluorescence was conducted to visualize the effects of ZNF408 mutants on cellular localization of ZNF408 in APRE-19 cells. Preliminary findings showed ZNF408 mutants may interact with different proteins as compared with wild-type ZNF408, suggesting these variants are causal. Further in vivo experiments using ZNF408 point mutation mouse models are necessary to better understand the effects of ZNF408 missense variant in Retinitis Pigmentosa pathology. This information will hopefully lead to better diagnostic approaches and potential therapeutic treatments for individuals present with mutations in this specific gene.

For more information: ftopham@eckerd.edu

Ruan Lab Website: https://www.theruanlabofrnametabolism.org/our-team 

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