Open AccessUrine mRNA to identify a novel pseudoexon causing dystrophinopathy
Author(s) -
Antoury Layal,
Hu Ningyan,
Darras Basil,
Wheeler Thurman M.
Publication year - 2019
Publication title -
annals of clinical and translational neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.824
H-Index - 42
ISSN - 2328-9503
DOI - 10.1002/acn3.777
Subject(s) - urine , muscle biopsy , medicine , rna , splice , messenger rna , digital polymerase chain reaction , complementary dna , biopsy , computational biology , gene , microbiology and biotechnology , pathology , genetics , biology , polymerase chain reaction
In muscular dystrophies, identification of pathogenic pseudoexons involves sequencing of the target gene cDNA derived from muscle mRNA. Here we use a urine “liquid biopsy,” droplet digital PCR, and sequencing of PCR products to identify a novel cryptic splice site in DMD intron 67 that causes dystrophinopathy. Pseudoexon inclusion is 35% in urine cells, 34% in urine extracellular RNA (exRNA), and 54% in muscle biopsy tissue, but absent in serum exRNA. Our results suggest that cryptic splice site use varies depending on the RNA source, and that urine RNA has the potential to substitute for muscle biopsies to identify DMD pseudoexons.