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Aberrant expression of PAX6 gene associated with classical aniridia: identification and functional characterization of novel noncoding mutations

Journal of Human Genetics volume 66, pages 333–338 (2021)Cite this article

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Abstract

The PAX6 is essential for ocular morphogenesis and is known to be highly sensitive to changes in gene expression, where neither over- nor under-expression ensures normal ocular development. Two unrelated probands with classical aniridia who were previously considered “PAX6-negative”, were studied by whole-genome sequencing. Through the use of multiple in silico deep learning-based algorithms, we identified two novel putative causal mutations, c.-133_-132del in the 5′ untranslated region (5′-UTR) and c.-52 + 5G>A in an intron upstream of the PAX6 gene. The luciferase activity was significantly increased and VAX2 binding was disrupted with the former 5′-UTR variant compared with wild-type sequence, which resulted in a striking overexpression of PAX6. The minigene assay showed that the c.-52 + 5G>A mutation caused defective splicing, which resulted in the formation of truncated transcripts.

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Acknowledgements

The authors would like to thank aniridia subjects and family members for their participation. We also thank to Dr K.A. Lukyanov and Dr Jong In Yook who provided us vectors.

Funding

This study was supported by a grant (no. 2019IL0366) from the Asan Institute for Life Sciences, Asan Medical Center, Seoul, Korea, a fund (2018-ER6902-00) by Research of Korea Centers for Disease Control and Prevention, and the National Research Foundation of Korea (NRF) grant funded by the Korean government [Ministry of Science and ICT; grant number: 2018R1C1B6002732].

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Author notes

  1. These authors contributed equally: Junwon Lee, Yoonjong Suh

  2. These authors jointly supervised this work: Jinu Han, Hyun Taek Lim

Authors and Affiliations

  1. Department of Ophthalmology, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea

    Junwon Lee, Yoonjong Suh, Han Jeong & Suk Ho Byeon

  2. Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, Korea

    Yoonjong Suh, Han Jeong & Suk Ho Byeon

  3. Medical Genetics Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea

    Gu-Hwan Kim

  4. Department of Ophthalmology, Gangnam Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea

    Jinu Han

  5. Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea

    Hyun Taek Lim

Authors
  1. Junwon Lee
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Contributions

JL, JH, and HTL wrote the paper and YS contributed to the paper. JL, YS, HJ, and SHB conceived the experiments, and designed strategies. GHK performed genotyping and segregation analysis. JL, SHB, JH, and HTL analyzed and interpreted data. JH performed the WGS analysis. All authors reviewed the paper.

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Correspondence to Jinu Han or Hyun Taek Lim.

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Lee, J., Suh, Y., Jeong, H. et al. Aberrant expression of PAX6 gene associated with classical aniridia: identification and functional characterization of novel noncoding mutations. J Hum Genet 66, 333–338 (2021). https://doi.org/10.1038/s10038-020-00829-2

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