Evaluation of both exonic and intronic variants for effects on RNA splicing allows for accurate assessment of the effectiveness of precision therapies

• DOI: 10.1371/journal.pgen.1009100
• Publication Date: 2020-10-21T17:37:22Z
• Authors: Anya T. Joynt; Taylor A. Evans; Matthew J. Pellicore; Emily F. Davis-Ma...; Melis A. Aksit; Alice C. Eastman; Shivani U. Patel; Kathleen C. Paul; Derek L. Osorio; Alyssa D. Bowling; Calvin U. Cotton; Karen S. Raraigh; Natalie E. West; Christian A. Merlo; Garry R. Cutting; Neeraj Sharma
• Keywords: Exon skipping

Elucidating the functional consequence of molecular defects underlying genetic diseases enables appropriate design therapeutic options. Treatment cystic fibrosis (CF) is an exemplar this paradigm as development CFTR modulator therapies has allowed for targeted and effective treatment individuals harboring specific variants. However, mechanism these drugs limits effectiveness to particular classes variants that allow production protein. Thus, assessment individual imperative proper assignment precision therapies. This particularly important when considering affect pre-mRNA splicing, thus limiting success existing protein-targeted Variants affecting splicing can occur throughout exons introns complexity process lends itself a variety outcomes, both at RNA protein levels, further complicating disease liability response. To investigate scope challenge, we evaluated downstream effects 52 naturally occurring (exonic = 15, intronic 37). Expression constructs containing select sequences complete exonic in cell line models protein-level on allele by basis. Characterization primary nasal epithelial cells obtained from splice corroborated vitro data. Notably, identified result missplicing lack response (e.g. c.2908G>A, c.523A>G), well respond modulators due presence residual normally spliced transcript c.4242+2T>C, c.3717+40A>G). Overall, our data reveals diverse outcomes amongst emphasizing need delineate RNA, protein, each variant order accurately assign