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Analysis of balanced reciprocal translocations in patients with subfertility using single-molecule optical mapping

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Abstract

Purpose

Approximately 1% of individuals who carry a balanced reciprocal translocation (BRT) are subfertile. Current karyotyping does not have the resolution to determine whether the breakpoints of the involved chromosomes perturb genes important for fertility. The aim of this study was to apply single-molecule optical mapping (SMOM) to patients presenting for IVF (in vitro fertilization) to ascertain whether the BRT disrupted any genes associated with normal fertility.

Methods

Nine subfertile patients with different BRTs were recruited for the study. Methyltransferase enzyme DLE1 was used to fluorescently label their genomic DNA samples at the recognition motif CTTAAG. The SMOM was performed on the Bionano platform, and long molecules aligned against the reference genome hg19 to identify the breakpoint regions. Mate-pair and PCR-Sanger sequencing were used to confirm the precise breakpoint sequences.

Results

Both breakpoint regions in each of the nine BRTs were finely mapped to small regions of approximately 10 Kb, and their positions were consistent with original cytogenetic banding patterns determined by karyotyping. In three BRTs, breakpoints disrupted genes known to be associated with male infertility, namely NUP155 and FNDC3A [46,XY,t(5;13)(p15;q22)], DPY19L1 [46,XY,t(1;7)(p36.3;p15), and BAI3 [46,XY,t(3;6)(p21;q16)].

Conclusions

The SMOM has potential clinical application as a rapid tool to screen patients with BRTs for underlying genetic causes of infertility and other diseases.

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Acknowledgments

The authors thank the patients who participated in this research study.

Funding

This work was supported by National Key Research and Development Program of China (2018YFC1003100).

Author information

Author notes

  1. Hui Wang, Zhengjun Jia and Aiping Mao contributed equally to this work.

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, PLA General Hospital, Beijing, 100853, China

    Hui Wang, Bing Xu, Shuling Wang, Sai Liu & Yuanqing Yao

  2. Prenatal Diagnosis Center of Hunan Province, Hunan Provincial Maternal and Child Health Care Hospital, Changsha, 410008, China

    Zhengjun Jia

  3. Berry Genomics Corporation, Beijing, 102200, China

    Aiping Mao, Haiman Zhang, Xiaojie Zhang, Tao Yu, Ting Mu, Mengnan Xu & David S. Cram

  4. The First Hospital of KunMing, Kunming, 650034, China

    Li Wang & Sai Liu

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  1. Hui Wang
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Contributions

H.W. and B.X.: study design and management and supervision of the study. A.M. and D.S.C.: study design, statistical analysis, and manuscript preparation. Z.J., A.M., H.Z., and T.Y.: experimental procedures and statistical analysis. X.Z. and T.M.: data analysis. S.W., L.W., and S.L.: supervision. M.X.: data analysis and manuscript preparation. Y.Y.: supervision and manuscript review.

Corresponding authors

Correspondence to David S. Cram or Yuanqing Yao.

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Conflict of interest

A.M., H.Z., X.Z., T.Y, T.M., M.X., and D.S.C. are employees of Berry Genomics Corporation. None of the authors holds any stocks or bonds in the company. The other authors declare that they have no conflict of interest.

Ethical approval

Ethical approval for performing this study was obtained from the Ethics Committee of the Chinese PLA General Hospital (Approval number S2013-092-02).

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Wang, H., Jia, Z., Mao, A. et al. Analysis of balanced reciprocal translocations in patients with subfertility using single-molecule optical mapping. J Assist Reprod Genet 37, 509–516 (2020). https://doi.org/10.1007/s10815-020-01702-z

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