Hui Wang1, Zhengjun Jia2, Aiping Mao3, Bing Xu1, Shuling Wang1, Li Wang4, Sai Liu1,4, Haiman Zhang3, Xiaojie Zhang3, Tao Yu3, Ting Mu3, Mengnan Xu3, David S Cram5, Yuanqing Yao6. 1. Department of Obstetrics and Gynecology, PLA General Hospital, Beijing, 100853, China. 2. Prenatal Diagnosis Center of Hunan Province, Hunan Provincial Maternal and Child Health Care Hospital, Changsha, 410008, China. 3. Berry Genomics Corporation, Beijing, 102200, China. 4. The First Hospital of KunMing, Kunming, 650034, China. 5. Berry Genomics Corporation, Beijing, 102200, China. david.cram@berrygenomics.com. 6. Department of Obstetrics and Gynecology, PLA General Hospital, Beijing, 100853, China. yqyao@126.com.
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.
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.
Authors: Isabelle Koscinski; Elias Elinati; Camille Fossard; Claire Redin; Jean Muller; Juan Velez de la Calle; Françoise Schmitt; Mariem Ben Khelifa; Pierre F Ray; Pierre Ray; Zaid Kilani; Christopher L R Barratt; Stéphane Viville Journal: Am J Hum Genet Date: 2011-03-11 Impact factor: 11.025
Authors: Michal Levy-Sakin; Steven Pastor; Yulia Mostovoy; Le Li; Alden K Y Leung; Jennifer McCaffrey; Eleanor Young; Ernest T Lam; Alex R Hastie; Karen H Y Wong; Claire Y L Chung; Walfred Ma; Justin Sibert; Ramakrishnan Rajagopalan; Nana Jin; Eugene Y C Chow; Catherine Chu; Annie Poon; Chin Lin; Ahmed Naguib; Wei-Ping Wang; Han Cao; Ting-Fung Chan; Kevin Y Yip; Ming Xiao; Pui-Yan Kwok Journal: Nat Commun Date: 2019-03-04 Impact factor: 14.919