Case Report: Minigene assays reveal a novel DNAAF6 intronic variant as the key etiology for primary ciliary dyskinesia - study

Sep 28, 2026

Primary ciliary dyskinesia (PCD) is a rare inherited disorder marked by impaired motility of motile cilia and is often linked with infertility. A family study addressed limited evidence for DNAAF6 intronic variants in non-classical splice regions. Whole-exome sequencing identified a novel hemizygous intronic variant in DNAAF6, confirmed by Sanger sequencing in an affected brother. Minigene splicing assays showed exon six skipping with a frameshift, and transmission electron microscopy demonstrated absent dynein arms, supporting flagellar immotility.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages8

Reading Time20 min

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Background

Case presentation

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TYPE Case Report PUBLISHED 18 December 2025 DOI 10.3389/fgene.2025.1692658 OPEN ACCESS EDITED BY Zheng Jin Tu, Cleveland Clinic, United States REVIEWED BY Doris Pinto-Escalante, Universidad Autónoma de Yucatán, Mexico Ao Ma, University of Science and Technology of China, China *CORRESPONDENCE Xuemei Yang, [renew_y@hotmail.com](renew_y@hotmail.com) RECEIVED 26 August 2025 REVISED 05 December 2025 ACCEPTED 08 December 2025 PUBLISHED 18 December 2025 CITATION Long Y, Li H, Yu H, Yi Q, Meng X, Wang X, Ren Q, Ding D, Li H, Zhang F, Qiu H and Yang X (2025) Case Report: Minigene assays reveal a novel DNAAF6 intronic variant as the key etiology for primary ciliary dyskinesia. Front. Genet. 16:1692658 . doi: 10.3389/fgene.2025.1692658 COPYRIGHT © 2025 Long, Li, Yu, Yi, Meng, Wang, Ren, Ding, Li, Zhang, Qiu and Yang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) . The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Case Report: Minigene assays reveal a novel DNAAF6 intronic variant as the key etiology for primary ciliary dyskinesia Yupeng Long 1, Huixian Li 2, Haoyue Yu 3, Qian Yi 3, Xiong Meng 3, Xi Wang 3, Qinqin Ren 3, Dongyan Ding 3, Haidong Li 3, Fenglan Zhang 2, Hao Qiu 2 and Xuemei Yang 3* 1Laboratory and Blood Transfusion Department of Jiangbei Campus, The First Affiliated Hospital of Army Medical University (The 958th hospital of Chinese People’s Liberation Army), Chongqing, China, 2Center for Clinical Genetics and Genomics, Dian Diagnostics Group Co., Ltd., Hangzhou, Zhejiang, China, 3Respiratory and Critical Care Medicine Department of Jiangbei Campus, The First Affiliated Hospital of Army Medical University (The 958th hospital of Chinese People’s Liberation Army), Chongqing, China Background: Primary ciliary dyskinesia (PCD), a rare hereditary disorder characterized by impaired ciliary motility, is frequently linked to infertility. Elucidating PCD’s genetic basis is critical for accurate diagnosis and clinical management. However, research on the pathogenicity of intronic variants in non-classical splice regions of DNAAF6—a newly identified PCD-associated gene in recent years—remains scarce. Case presentation: A proband with primary ciliary dyskinesia (PCD) and male infertility underwent whole exome sequencing (WES), revealing a novel hemizygous intronic variant in the DNAAF6 gene (NM_173494. 2: c.515 + 3_ 515+6del, Clinvar Variation ID:4075425) . Sanger sequencing confirmed this variant in his affected brother. A minigene splicing assay showed that the variant caused exon six skipping, leading to a frameshift. Transmission electron microscopy (TEM) analysis of sperm flagella indicated absent outer and inner dynein arms, resulting in flagellar immotility. Conclusion: This study identifies and confirms a novel pathogenic intronic variant of DNAAF6 in a Chinese PCD family, expanding the DNAAF6 mutational spectrum. It also emphasizes the critical role of minigene-based functional validation in interpreting variants of uncertain significance (VUS) . KEYWORDS case report, DNAAF6 gene, male infertility, minigene, primary ciliary dyskinesia 1 Introduction Primary ciliary dyskinesia (PCD) represents a rare, inherited disorder specifically affecting motile cilia. This condition is precipitated by genetic variations that give rise to abnormalities in both the structure and function of cilia. Consequentially, these aberrations lead to disorders in the movement of ciliary epithelium within multiple organs. As a result, organs harboring ciliary tissue experience dysfunction, thereby posing a significant threat to human health. T

FAQs

What genetic change was identified in the DNAAF6 gene in this case report?

Whole-exome sequencing found a novel hemizygous intronic variant in DNAAF6, described as NM_173494.2: c.515+3_515+6del. Sanger sequencing confirmed the variant in the affected brother.

How did the minigene splicing assay clarify the variant’s effect?

The minigene assay showed that the variant caused exon six skipping, leading to a frameshift, supporting pathogenic impact on mRNA splicing and protein coding.

What did transmission electron microscopy reveal about sperm flagella?

Transmission electron microscopy of sperm flagella indicated absent outer and inner dynein arms, consistent with flagellar immotility.

Sep 28, 2026

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