1. Multiplexed Functional Assessment of Genetic Variants in CARD11.
- Author
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Meitlis I, Allenspach EJ, Bauman BM, Phan IQ, Dabbah G, Schmitt EG, Camp ND, Torgerson TR, Nickerson DA, Bamshad MJ, Hagin D, Luthers CR, Stinson JR, Gray J, Lundgren I, Church JA, Butte MJ, Jordan MB, Aceves SS, Schwartz DM, Milner JD, Schuval S, Skoda-Smith S, Cooper MA, Starita LM, Rawlings DJ, Snow AL, and James RG
- Subjects
- Adenine analogs & derivatives, Adenine pharmacology, B-Cell CLL-Lymphoma 10 Protein genetics, B-Lymphocytes cytology, Cell Line, Diploidy, Exons, Genes, Dominant, Humans, Jurkat Cells, Lymphoma genetics, NF-kappa B p50 Subunit genetics, Piperidines pharmacology, Polymorphism, Single Nucleotide, Primary Immunodeficiency Diseases genetics, Sensitivity and Specificity, CARD Signaling Adaptor Proteins genetics, Genetic Variation, Guanylate Cyclase genetics, Immunologic Deficiency Syndromes genetics
- Abstract
Genetic testing has increased the number of variants identified in disease genes, but the diagnostic utility is limited by lack of understanding variant function. CARD11 encodes an adaptor protein that expresses dominant-negative and gain-of-function variants associated with distinct immunodeficiencies. Here, we used a "cloning-free" saturation genome editing approach in a diploid cell line to simultaneously score 2,542 variants for decreased or increased function in the region of CARD11 associated with immunodeficiency. We also described an exon-skipping mechanism for CARD11 dominant-negative activity. The classification of reported clinical variants was sensitive (94.6%) and specific (88.9%), which rendered the data immediately useful for interpretation of seven coding and splicing variants implicated in immunodeficiency found in our clinic. This approach is generalizable for variant interpretation in many other clinically actionable genes, in any relevant cell type., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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