例如:"lncRNA", "apoptosis", "WRKY"

A CRISPR and high-content imaging assay compliant with ACMG/AMP guidelines for clinical variant interpretation in ciliopathies.

Hum Genet. 2021 Apr;140(4):593-607. Epub 2020 Oct 23
Liliya Nazlamova 1 , N Simon Thomas 2 , Man-Kim Cheung 3 , Jelmer Legebeke 1 , Jenny Lord 4 , Reuben J Pengelly 4 , William J Tapper 1 , Gabrielle Wheway 5
Liliya Nazlamova 1 , N Simon Thomas 2 , Man-Kim Cheung 3 , Jelmer Legebeke 1 , Jenny Lord 4 , Reuben J Pengelly 4 , William J Tapper 1 , Gabrielle Wheway 5
+ et al

[No authors listed]

Author information
  • 1 Faculty of Medicine, Human Development and Health, University of Southampton, Southampton, UK.
  • 2 Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, UK.
  • 3 Centre for Research in Biosciences, University of the West of England, Bristol, Bristol, UK.
  • 4 University Hospital Southampton NHS Foundation Trust, Southampton, UK.
  • 5 University Hospital Southampton NHS Foundation Trust, Southampton, UK. G.Wheway@soton.ac.uk.

摘要


Ciliopathies are a broad range of inherited developmental and degenerative diseases associated with structural or functional defects in motile or primary non-motile cilia. There are around 200 known ciliopathy disease genes and whilst genetic testing can provide an accurate diagnosis, 24-60% of ciliopathy patients who undergo genetic testing do not receive a genetic diagnosis. This is partly because following current guidelines from the American College of Medical Genetics and the Association for Molecular Pathology, it is difficult to provide a confident clinical diagnosis of disease caused by missense or non-coding variants, which account for more than one-third of cases of disease. Mutations in PRPF31 are the second most common cause of the degenerative retinal ciliopathy autosomal dominant retinitis pigmentosa. Here, we present a high-throughput high-content imaging assay providing quantitative measure of effect of missense variants in PRPF31 which meets the recently published criteria for a baseline standard in vitro test for clinical variant interpretation. This assay utilizes a new PRPF31+/- human retinal cell line generated using CRISPR gene editing to provide a stable cell line with significantly fewer cilia in which novel missense variants are expressed and characterised. We show that high-content imaging of cells expressing missense variants in a ciliopathy gene on a null background can allow characterisation of variants according to the cilia phenotype. We hope that this will be a useful tool for clinical characterisation of PRPF31 variants of uncertain significance, and can be extended to variant classification in other ciliopathies.