Uncovering Pre‐messenger RNA Splicing Mechanisms in Retinitis Pigmentosa

Abstract

Disruptions of pre‐messenger RNA splicing is a common mechanism found in human diseases, like autosomal dominant Retinitis Pigmentosa (adRP). adRP affects ~1 in 4,000 people and is associated with the breakdown and loss of cells in the peripheral retina. Genetic analyses of patients harboring adRP have linked eight proteins that are associated with the spliceosome to adRP. Currently, there are two working hypotheses for how mutations in these splicing proteins contribute to adRP. First, splicing defects are occurring ubiquitously in all tissues but are more noticeable in the retinal tissue. Alternatively, that these splicing defects are strictly happening in the retina. This study focuses on understanding how mutations in one of these splicing proteins, PRPF31, impact splicing and cellular growth. In this study, we use a mouse retinal ganglion precursor‐like cell line model to identify various mRNAs effects of adRP mutants splicing function. We are examining six prpf31 mutants adRP mutants selected from patients with adRP. Expression constructs have been prepared for wildtype and our adRP mutants and successfully transfected into the mouse cell lines. We have also established a splicing assay for our mouse photoreceptor cell studies utilizing Pladienolide B, a splicing inhibitor as a model, and will examine the PRPF31 mutants using this assay. Our ongoing work will be described.