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Identification of RPGR ORF15 mutation for X-linked retinitis pigmentosa in a large Chinese family and in vitro correction with prime editor

Gene Therapy volume 30, pages 160–166 (2023)Cite this article

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Abstract

X-linked retinitis pigmentosa (XLRP) is the most severe form of Retinitis Pigmentosa (RP) and one of the leading causes of blindness in the world. Currently, there is no effective treatment for RP. In the present study, we recruited a XLRP family and identified a 4 bp deletion mutation (c. 2234_2237del) in RPGR ORF15 with Sanger sequencing, which was located in the exact same region as the missing XES (X chromosome exome sequencing) coverage. Then, we generated cell lines harboring the identified mutation and corrected it via enhanced prime editing system (ePE). Collectively, Sanger sequencing identified a pathogenic mutation in RPGR ORF15 for XLRP which was corrected with ePE. This study provides a valuable insight for genetic counseling of the afflicted family members and prenatal diagnosis, also paves a way for applying prime editing based gene therapy in those patients.

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Fig. 1: Clinical data for X-linked retinitis pigmentosa patient.
Fig. 2: Schematic of gene correction in HEK293-mRO cell line.
Fig. 3: PE2 and ePE-mediated gene correction in HEK293-mRO cells.

Data availability

The corresponding author declares that the data are available if requested. The deep sequencing data are available at the NCBI Sequence Read Archive (SRA) under BioProject PRJNA#573504 and PRJNA783078 (SRA accession number SRR17023438 and SRP17023439, sample accession numbers, SAMN23416124).

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Acknowledgements

We thank the families for their participation in this project. We are indebted to Dr. Jinyu Wu (Institute of Genomic Medicine, Wenzhou Medical University) for bioinformatics analysis and Prof.Peter Reinach for comments on the manuscript. This work was supported by grants from National Natural Science Foundation of China (81201181), Science Technology project of Zhejiang Province (2017C37176), and Project of State Key Laboratory of Ophthalmology, Optometry and Visual Science, Wenzhou Medical University (J02–20190201) and Wenzhou City Grant (H20210011).

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Author notes

  1. These authors contributed equally: Xiujuan Lv, Zheng Zheng.

Authors and Affiliations

  1. School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, State Key Laboratory and Key Laboratory of Vision Science, Ministry of Health and Zhejiang Provincial Key Laboratory of Ophthalmology and Optometry, Wenzhou, Zhejiang, 325027, China

    Xiujuan Lv, Zheng Zheng, Xiao Zhi, Jineng Lv, Yue Zhou, Binrong Wu, Sixiu Liu, Jinling Xu, Jia Qu, Dan Xu & Feng Gu

  2. College of Engineering, Boston University, Boston, MA, USA

    Yilin Zhou

  3. Department of Ophthalmology, Beijing Children’s Hospital, Capital Medical University, Beijing, 100054, China

    Wei Shi

  4. Henan Eye Hospital, Henan Eye Institute, Henan Provincial People’s Hospital and People’s Hospital of Zhengzhou University and People’s Hospital of Henan University, Zhengzhou, Henan, China

    Zongming Song

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Contributions

XL and FG conceived and designed the study experiments. ZZ and XZ identified the mutation. XL, ZZ, DX, and FG wrote and edited manuscript. XL, YZ, JL, YZ, BW, SL, WS performed the experiments. ZS, JX, JQ, and FG contributed to data analysis, data interpretation. XL, DX, and FG revised the manuscript. All authors have read and approved the final manuscript, and therefore, have full access to all data in this study and take responsibility for the integrity and security of the data.

Corresponding authors

Correspondence to Dan Xu or Feng Gu.

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The authors declare no competing interests.

Ethics approval and consent to participate

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008 (5). Informed consent was obtained from all patients for being included in the study. Additional informed consent was obtained from all patients for which identifying information is included in this article.

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Lv, X., Zheng, Z., Zhi, X. et al. Identification of RPGR ORF15 mutation for X-linked retinitis pigmentosa in a large Chinese family and in vitro correction with prime editor. Gene Ther 30, 160–166 (2023). https://doi.org/10.1038/s41434-022-00352-3

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