Abstract
Embryo development can be affected by genetic material and growth environment. Embryo
genetic abnormalities, which originate from paternal, maternal or embryonic genome, may
increase the risk of implantation failures and human infertility. Next generation sequencing
technologies have been applied in many fields including reproductive and developmental
sciences. The characters of NGS, high throughputs and accuracy at base-pair resolution, can help
us to know not only the key molecular events during early embryo development, but also the
factors correlated with infertility and miscarriage. In Part I of this thesis, single cells were
isolated from four groups of developing embryos，two groups of androgenetic and two groups
of parthenogenetic embryos at four embryonic stages. Their genome and transcriptome were
investigated using single cell whole genome and transcriptome sequencing based on NGS
technologies. Our results implied that parent-of-origin effects played a predominant role after the
major wave of embryonic genome activation (EGA) during embryo development. In Part II and
III of this thesis, low-pass whole genome sequencing was successfully used to detect seemingly
balanced chromosomal abnormalities (BCAs) in recurrent miscarriage population and normal
population from 1000 genomes project. Comparing with traditional chromosome analysis, lowpass
WGS also can identify cryptic rearrangements and provided important information for
complex chromosome rearrangements in blind. Additionally, combined with RNA expression
analysis, it shown that the expression levels of some genes in breakpoints areas were not
changed, which indicated that the clinical interpretation for balanced chromosome abnormality
(BCA) would be better if combined with RNA expression analysis. These results show that lowpass
WGS is a highly cost-effective approach to detect BCA events. Taken together, NGS has
wide applications in reproductive and developmental sciences, from basic research in embryo
development to clinical applications for abnormal chromosome identification.
genetic abnormalities, which originate from paternal, maternal or embryonic genome, may
increase the risk of implantation failures and human infertility. Next generation sequencing
technologies have been applied in many fields including reproductive and developmental
sciences. The characters of NGS, high throughputs and accuracy at base-pair resolution, can help
us to know not only the key molecular events during early embryo development, but also the
factors correlated with infertility and miscarriage. In Part I of this thesis, single cells were
isolated from four groups of developing embryos，two groups of androgenetic and two groups
of parthenogenetic embryos at four embryonic stages. Their genome and transcriptome were
investigated using single cell whole genome and transcriptome sequencing based on NGS
technologies. Our results implied that parent-of-origin effects played a predominant role after the
major wave of embryonic genome activation (EGA) during embryo development. In Part II and
III of this thesis, low-pass whole genome sequencing was successfully used to detect seemingly
balanced chromosomal abnormalities (BCAs) in recurrent miscarriage population and normal
population from 1000 genomes project. Comparing with traditional chromosome analysis, lowpass
WGS also can identify cryptic rearrangements and provided important information for
complex chromosome rearrangements in blind. Additionally, combined with RNA expression
analysis, it shown that the expression levels of some genes in breakpoints areas were not
changed, which indicated that the clinical interpretation for balanced chromosome abnormality
(BCA) would be better if combined with RNA expression analysis. These results show that lowpass
WGS is a highly cost-effective approach to detect BCA events. Taken together, NGS has
wide applications in reproductive and developmental sciences, from basic research in embryo
development to clinical applications for abnormal chromosome identification.
| Originalsprog | Engelsk |
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| Forlag | Department of Biology, Faculty of Science, University of Copenhagen |
|---|---|
| Status | Udgivet - 2018 |