Project description:Mouse Wdr74-knockout embryos were generated using the CRISPR-Cas9 system. Embryos from the 4-cell stage to the morula stage were collected and treated with acidic Tyrode's solution to remove the zona pellucida. We used 50 embryos per sample at each developmental stage and identified the proteome of mouse Wdr74-knockout embryos by label-free quantification.
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:Data showing the late 2-cell-stage, control embryos (Imp2♀+/♂+) and Imp2-knockout embryos (Imp2♀−/♂+) for HPLC MS/MS analysis. 3 replicates were performed using 330 embryos per group.
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:Mouse oocytes and embryos from the germinal vesicle (GV) stage to the blastocyst stage were collected and treated with acidic Tyrode’s solution to remove the zona pellucida. We used 42.5 oocytes or embryos per sample at each developmental stage and identified the proteome during mouse pre-implantation development by label-free quantification.
