Predicting the pathogenicity of RPE65 mutations

A. R. Philpa, M. Jin, S. Li, E. I. Schindler, A. Iannaccone, B. L. Lam, R. G. Weleber, G. A. Fishman, S. G. Jacobson, R. F. Mullins, Gabriel H. Travis, Edwin M. Stone

Research output: Contribution to journalArticlepeer-review

42 Scopus citations


To assist in distinguishing disease-causing mutations from nonpathogenic polymorphisms, we developed an objective algorithm to calculate an "estimate of pathogenic probability" (EPP) based on the prevalence of a specific variation, its segregation within families, and its predicted effects on protein structure. Eleven missense variations in the RPE65 gene were evaluated in patients with Leber congenital amaurosis (LCA) using the EPP algorithm. The accuracy of the EPP algorithm was evaluated using a cell-culture assay of RPE65-isomerase activity The variations were engineered into plasmids containing a human RPE65 cDNA and the retinoid isomerase activity of each variant was determined in cultured cells. The EPP algorithm predicted eight substitution mutations to be disease-causing variants. The isomerase catalytic activities of these RPE65 variants were all less than 6% of wild-type. In contrast, the EPP algorithm predicted the other three substitutions to be non-disease-causing, with isomerase activities of 68%, 127%, and 110% of wild-type, respectively. We observed complete concordance between the predicted pathogenicities of missense variations in the RPE65 gene and retinoid isomerase activities measured in a functional assay. These results suggest that the EPP algorithm may be useful to evaluate the pathogenicity of missense variations in other disease genes where functional assays are not available.

Original languageEnglish (US)
Pages (from-to)1183-1188
Number of pages6
JournalHuman mutation
Issue number8
StatePublished - Aug 2009


  • Leber congenital amaurosis
  • RPE65
  • Retinoid

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)


Dive into the research topics of 'Predicting the pathogenicity of RPE65 mutations'. Together they form a unique fingerprint.

Cite this