Project description:We used single-embryo sequencing to characterize early zygotic gene expression in Drosophila. The data provide the first continuous timeline of transcript levels during early development (≤ 3 hours) in Drosophila melanogaster.
Project description:To identify Aub function in mRNA regulation in the Drosophila embryo, we have performed mass spectrometry analysis of Aub interactors, following immunoprecipitation of GFP-Aub in 0-2 hour-embryos. Immunoprecipitation of GFP alone was used as negative control. Because Aub accumulates at high levels in the germ plasm, GFP-Aub immunoprecipitation was also performed in oskar mutant embryos that do not assemble the germ plasm. Proteins coprecipitating with GFP-Aub were similar in wild-type and oskar mutant embryos. Translation factors were enriched among proteins coprecipitating with Aub.
Project description:The gfzf gene is essential for viability and it has been identified in several genetic screenings pointed out its importance in different cellular processes. However, its molecular function has not been extensively studied so far. Here we analyse the GFZF protein interactome in Drosophila melanogaster embryos. The GFZF protein was subjected to immunoprecipitation with an antibody against the endogenous protein and the interactome was analysed by mass spectrometry. The samples are named as follows: antibody used _ batch _ MS ID _ date of MS run
Project description:BACKGROUND:
Preimplantation genetic diagnosis (PGD) enables profiling of embryos for genetic disorders prior to implantation. The majority of PGD testing is restricted in the scope of variants assayed or by the availability of extended family members. While recent advances in single cell sequencing show promise, they remain limited by bias in DNA amplification and the rapid turnaround time (<36 h) required for fresh embryo transfer. Here, we describe and validate a method for inferring the inherited whole genome sequence of an embryo for preimplantation genetic diagnosis (PGD).
METHODS:
We combine haplotype-resolved, parental genome sequencing with rapid embryo genotyping to predict the whole genome sequence of a day-5 human embryo in a couple at risk of transmitting alpha-thalassemia.
RESULTS:
Inheritance was predicted at approximately 3 million paternally and/or maternally heterozygous sites with greater than 99% accuracy. Furthermore, we successfully phase and predict the transmission of an HBA1/HBA2 deletion from each parent.
CONCLUSIONS:
Our results suggest that preimplantation whole genome prediction may facilitate the comprehensive diagnosis of diseases with a known genetic basis in embryos.
Project description:The use of in vitro fertilization (IVF) has revolutionized the treatment of infertility and is now responsible for 1-5% of all births in industrialized countries. During IVF, it is typical for patients to generate multiple embryos. However, only a small proportion of them possess the genetic and metabolic requirements needed in order to produce a healthy pregnancy. The identification of the embryo with the greatest developmental capacity represents a major challenge for fertility clinics. Current methods for the assessment of embryo competence are proven inefficient and the inadvertent transfer of non-viable embryos is the principal reason why most IVF treatments (approximately two-thirds) end in failure. In this study, we investigate how the application of proteomic measurements could improve success rates in clinical embryology. We describe a procedure that allows the identification and quantification of proteins of embryonic origin, present in attomole concentrations in the blastocoel, the enclosed fluid filled cavity that forms within five-day old human embryos. By using targeted proteomics, we demonstrate the feasibility of quantifying multiple proteins in samples derived from single blastocoels, and that such measurements correlate with aspects of embryo viability, such as chromosomal (ploidy) status. This study illustrates the potential of high-sensitivity proteomics to measure clinically relevant biomarkers in minute samples and, more specifically, suggests that key aspects of embryo competence could be measured using a proteomic-based strategy, with negligible risk of harm to the living embryo. Our work paves the way for the development of “next-generation” embryo competence assessment strategies, based on functional proteomics.
Project description:Global untargeted metabolomics study to analyse culture media extracted from an innovative microfluidic device or traditional microdrops in presence or absence of murine embryos to investigate PDMS-release of biomolecules and embryo metabolic activity.
Project description:The aim of the study was to compare the effects of maternal age on embryos in horses. For that, 11 embryos were collected at 8 days post ovulation (expanded blastocyst stage) from 2 groups of multiparous mares of different age: 5 from young mares (<10 years old) and 6 from old mares (10-17 years old). Embryos collected were cut in 2 parts: one hemi-embryo containing pure trophoblast and the other one containing trophoblast + inner cell mass. ARN extractions were performed for all samples using PicoPure RNA isolation kit (Applied Biosystem). Five nanograms of both parts of each embryo were sequenced in paired-end with a length of 30-50bp separately using Illumina NextSeq 500 High. Data were trimmed using Cutadapt. Sequences with less than 10bp were removed.
Project description:The aim of the global study was to compare the effects of maternal age and parity on gene expression of equine embryos. For that, 17 embryos were collected at 8 days post ovulation (expanded blastocyst stage) from 3 groups of young or old nulliparous or multiparous mares: 6 from young nulliparous mares (5 or 6 year-old, never foaled before), 5 from young multiparous mares (6 year-old and foaled at least once) and 6 from old multiparous (10-16 year-old and foaled at least once). Embryos collected were cut in 2 parts: one hemi-embryo containing pure trophoblast and the other one containing trophoblast + inner cell mass. ARN extractions were performed for all samples using PicoPure RNA isolation kit (Applied Biosystem). Five nanograms of both parts of each embryo were sequenced in paired-end with a length of 30-50bp separately using Illumina NextSeq 500 High. Data were trimmed using Cutadapt. Sequences with less than 10bp were removed. Data for young and old multiparous mares had been already published in [accession: GSE162893; https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE162893]. Only data for embryos collected from young nulliparous mares are deposited here
Project description:Comprehensive quantitative proteomic study of human pre-implantation embryo stages reveal dynamic proteome landscape from M2, 8-cell and blastocyst stage, and during trophoblast stem cell (TS) differentiation. Identified key factors in early human embryos and lineage-specific trophoblast proteome profiles, correlated with transcriptomic analyses. This direct proteomic analysis provides a comprehensive analysis of the dynamic protein expression in human embryos during pre-implantation development and a powerful resource to enable further mechanistic studies on human trophoblast development and function.