title: Reciprocal Translocation Carrier Diagnosis in Preimplantation Human Embryos.
abstract: Preimplantation genetic diagnosis (PGD) is widely applied in reciprocal translocation carriers to increase the chance for a successful live birth. However, reciprocal translocation carrier embryos were seldom discriminated from the normal ones mainly due to the technique restriction. Here we established a clinical applicable approach to identify precise breakpoint of reciprocal translocation and to further distinguish normal embryos in PGD. In the preclinical phase, rearrangement breakpoints and adjacent single nucleotide polymorphisms (SNPs) were characterized by next-generation sequencing following microdissecting junction region (MicroSeq) from 8 reciprocal translocation carriers. Junction-spanning PCR and sequencing further discovered precise breakpoints. The precise breakpoints were identified in 7/8 patients and we revealed that translocations in 6 patients caused 9 gene disruptions. In the clinical phase of embryo analysis, informative SNPs were chosen for linkage analyses combined with PCR analysis of the breakpoints to identify the carrier embryos. From 15 blastocysts diagnosed to be chromosomal balanced, 13 blastocysts were identified to be carriers and 2 to be normal. Late prenatal diagnoses for five carriers and one normal fetus confirmed the carrier diagnosis results. Our results suggest that MicroSeq can accurately evaluate the genetic risk of translocation carriers and carrier screen is possible in later PGD treatment.
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China; National Engineering and Research Center of Human Stem Cells, Changsha 410013, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Hu
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China.
authors.forename: Dehua
authors.initials: D
authors.lastname: Cheng
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Gong
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China.
authors.forename: Changfu
authors.initials: C
authors.lastname: Lu
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China; National Engineering and Research Center of Human Stem Cells, Changsha 410013, China.
authors.forename: Yueqiu
authors.initials: Y
authors.lastname: Tan
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China.
authors.forename: Keli
authors.initials: K
authors.lastname: Luo
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China.
authors.forename: Xianhong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China.
authors.forename: Wenbing
authors.initials: W
authors.lastname: He
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cells, Changsha 410013, China.
authors.forename: Pingyuan
authors.initials: P
authors.lastname: Xie
authors.affiliation: Peking Jabrehoo Med Tech., Ltd., Beijing 100089, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Feng
authors.affiliation: Peking Jabrehoo Med Tech., Ltd., Beijing 100089, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Yang
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China; National Engineering and Research Center of Human Stem Cells, Changsha 410013, China.
authors.forename: Guangxiu
authors.initials: G
authors.lastname: Lu
authors.affiliation: Institute of Reproduction and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410008, China; National Engineering and Research Center of Human Stem Cells, Changsha 410013, China. Electronic address: linggf@hotmail.com.
authors.forename: Ge
authors.initials: G
authors.lastname: Lin
mesh_terms: D001755:Blastocyst / Q000378:metabolism; D056905:Chromosome Breakpoints; D002874:Chromosome Mapping; D005260:Female; D005307:Fertilization in Vitro; D006580:Genetic Carrier Screening / Q000379:methods; D006579:Heterozygote; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D020641:Polymorphism, Single Nucleotide; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods; D014178:Translocation, Genetic
keywords: Breakpoint; Preimplantation genetic diagnosis; Reciprocal translocation; Single nucleotide polymorphism