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Functional evaluation of rare variants in complement factor I using a minigene assay
Journal article   Open access   Peer reviewed

Functional evaluation of rare variants in complement factor I using a minigene assay

Cobey J. H. Donelson, Nicolo Ghiringhelli Borsa, Amanda O. Taylor, Richard J. H. Smith and Yuzhou Zhang
Frontiers in immunology, Vol.15, 1446081
08/01/2024
DOI: 10.3389/fimmu.2024.1446081
PMCID: PMC11374653
PMID: 39238643
url
https://doi.org/10.3389/fimmu.2024.1446081View
Published (Version of record) Open Access

Abstract

The regulatory serine protease, complement factor I (FI), in conjunction with one of its cofactors (FH, C4BP, MCP, or CR1), plays an essential role in controlling complement activity through inactivation of C3b and C4b. The functional impact by missense variants in the CFI gene, particularly those with minor allele frequencies of 0.01% to 0.1%, is infrequently studied. As such, these variants are typically classified as variants of uncertain significance (VUS) when they are identified by clinical testing. Herein, we utilized a minigene splicing assay to assess the functional impact of 36 ultra-rare variants of CFI. These variants were selected based on their minor allele frequencies (MAF) and their association with low-normal FI levels. Four variants lead to aberrant splicing–one 5’ consensus splice site (NM_000204.5: c.1429G>C, p.Asp477His) and three exonic changes (c.355G>A, p.Gly119Arg; c.472G>A, p.Gly158Arg; and c.950G>A, p.Arg317Gln)–enabling their reclassification to likely pathogenic (LP) or pathogenic (P) based on ACMG guidelines. These findings underscore the value of functional assays, such as the minigene assay, in assessing the clinical relevance of rare variants in CFI.
alternative pathway complement complement factor I complement-mediated diseases haploinsufficiency RNA splicing

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