Identification of Cis-regulatory Elements Essential for Oocyte Maturation by Single-cell Parallel Sequencing on Human Oocytes
Refereed conference paper presented and published in conference proceedings


摘要Oocyte quality is critical for fertility, implantation and embryonic development. Currently, morphological assessment is widely clinically applied in predicting oocyte quality and embryo developmental potential. However, several studies have reported that the above method is limited and cannot accurately predict the success rate of assisted reproduction technology (ART). Thus, molecular assessment based on genetics and epigenetics signatures would be an alternative strategy. So far, transcriptome or methylome profiles of single human oocyte have been generated, separately.
In this study, we aimed to generate the simultaneous transcriptome and methylome landscapes from a single human oocyte by single-cell parallel methylome and transcriptome sequencing (scM&T-seq) and identified oocyte-specific cis-regulation elements. We performed scM&T-seq on two oocytes donated from a 32-years-old in vitro fertilization (IVF) patient. A total of 10,317 genes were shared in two oocytes, and they showed the high correlation of transcriptome (r=0.984, Pearson’s correlation test). For methylome profiles, reduced representation bisulfite sequencing (RRBS) was performed. In two samples, we detected average 343,196 CpG sites (222,335 and 464,047, separately) and two samples shared 3,450 CpG islands. The correlation coefficient of methylation level between two oocytes was up to 0.870 in 1000-bp bin size. A positive correlation between gene expression and methylation level in gene body, but not in promoter region, was observed. Furthermore, we obtained 72 common genes with highly expression level and low methylation level in promoters in two oocytes. Based on FANTOM5, Hocomoco (v11) and InWeb_InBioMap datasets, we identified 54 pairs of transcription factors (TF)-TF interaction in enhancer and promoter regions and total 29 genes were involved. Those genes, such as DCP1A, TPT1 and SCO2, have been reported to be closely associated with oocyte maturation, maintenance or aging.
In summary, we are the first to perform scM&T-seq on a single human oocyte. We demonstrated the two single oocytes from the same donor exhibited a positive correlation between gene expression and methylation level on gene body region. Importantly, to our knowledge, our study is the first to identify several novel cis-regulatory elements which are believed to regulate the transcription of the genes essential for oocyte maturation and quality. Our findings could make better understanding of transcriptional regulatory mechanism in oocyte and will provide a basis of developing the molecular measurement of oocyte quality.
著者QIAN Yan, LIAO Jinyue, CHI Ling, KONG Wing Shan Grace, LEUNG Tak Yeung, YIP Yuk-Lap Kevin, CHOW King Lau, LI Tin Chiu, CHAN Wai-Yee, LEE Tin-Lap
會議名稱ACMG Annual Clinical Genetics Meeting

上次更新時間 2018-05-06 於 10:48