Project description:By combining Nkx2.5-Cre x diphtheria toxin ablation (DTA) and interspecies blastocyst complementation we have generated rat hearts into mouse embryos at E10.5. These hearts have been compared by single cell-RNA sequencing with the hearts of stage matched E11.5 rat embryo. All heart specific cell types were found in complemented chimeras, and they showed high correlation with the control. This suggests that the mechanisms of formation of the heart are compatible between mouse and rat, and that all cardiac cell types are specified correctly in the rat-to-mouse chimeras. However, global gene expression comparison identified some genes differentially expressed between complemented chimeras and control. Notably, the most relevant transcriptomic differences between rPSCs-derived hearts and rat embryonic hearts were associated with metabolism, specifically, with a decrease in oxidative phosphorylation and an increase in glycolysis processes in the rPSCs-derived cells of chimeras. Additionally, vascular endothelial cells of the complemented chimeras augmented the expression of genes related to vasculogenesis and vascular development. These changes are suggestive of a low level of oxygen in the embryo.
Project description:Chimeric embryos were generated to investigate the effect of Mixl1 knockout in mouse embryos by single-cell RNA-sequencing. Mixl1 is an essential transcription factor for mesendoderm development. Chimeric embryos contain tissue that is Mixl1+/+, which prevents global developmental failures. Embryos were generated by blastocyst injection of tdTomato-labelled, Mixl1-/- mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.
Project description:Chimeric embryos were generated to investigate the effect of T knockout in mouse embryos by single-cell RNA-sequencing. T is an essential transcription factor for axial embryonic patterning. Chimeric embryos contain tissue that is T+/+, which prevents global developmental failures. Embryos were generated by blastocyst injection of tdTomato-labelled, Tal1-/- mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.
Project description:We performed single cell RNA-seq to quantify gene expression in forebrain tissue from the wild-type mouse, rat, and chimeric mouse. Single cell data from WT mouse or rat was used to perform integrative analysis for identifying rat cells and mouse cells in the chimaeras, respectively.
Project description:Chimeric embryos were generated to investigate the effect of Tal1 knockout in mouse embryos by single-cell RNA-sequencing. Tal1 is an essential transcription factor for the formation of the embryonic blood. Embryo chimerism permits the analysis of the effects of Tal1 knockout without the confounding effects of the absence of embryonic blood, which results in global developmental failures. Embryos were generated by blastocyst injection of tdTomato-labelled, Tal1-/- mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.
Project description:Chimeric mouse embryos were generated as a control experiment to understand the contribution of host vs. injected cells to the developing embryo. Embryos were generated by blastocyst injection of tdTomato-labelled mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.
Project description:This dataset contains additional data performed as in the experiment stored at accession E-MTAB-7324. Chimeric mouse embryos were generated as a control experiment to understand the contribution of host vs. injected cells to the developing embryo. Embryos were generated by blastocyst injection of tdTomato-labelled mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.
Project description:Chimeric embryos were generated to investigate the effect of Tal1 knockout in mouse embryos by single-cell RNA-sequencing. Tal1 is an essential transcription factor for the formation of the embryonic blood. Embryo chimerism permits the analysis of the effects of Tal1 knockout without the confounding effects of the absence of embryonic blood, which results in global developmental failures. Embryos were generated by blastocyst injection of tdTomato-labelled, Tal1-/- mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.
Project description:we report the application of single cell RNAseq analysis of mouse lung from E20.5 Nkx2-1-/- rat (mutant), and mouse-rat chimeric lung from E20.5 wt rat (control).