Cell-based analysis of CAD variants identifies individuals likely to benefit from uridine therapy.

ABSTRACT:

Purpose

Pathogenic autosomal recessive variants in CAD, encoding the multienzymatic protein initiating pyrimidine de novo biosynthesis, cause a severe inborn metabolic disorder treatable with a dietary supplement of uridine. This condition is difficult to diagnose given the large size of CAD with over 1000 missense variants and the nonspecific clinical presentation. We aimed to develop a reliable and discerning assay to assess the pathogenicity of CAD variants and to select affected individuals that might benefit from uridine therapy.

Methods

Using CRISPR/Cas9, we generated a human CAD-knockout cell line that requires uridine supplements for survival. Transient transfection of the knockout cells with recombinant CAD restores growth in absence of uridine. This system determin

SUBMITTER: Del Cano-Ochoa F 

PROVIDER: S-EPMC7521996 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature