Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse E8.5 wild-type and 3 E8.5 Tcof1+/- littermate embryos (Treacher Collins Syndrome)

ABSTRACT: The object of this study was to identify genes transcriptionally upregulated and downregulated in response to Tcof1 haploin-sufficiency during mouse embryogensis Experiment Overall Design: Total RNA was extracted from 3 E8.5 wild-type and 3 E8.5 Tcof1+/- littermate embryos using the RNeasy Mini Protocol for Isolation of Total RNA from Animal Tissues (Qiagen) according to the manufacturerâs protocol. RNA quality and quantity was determined using the Bioanalyser. To generate targets for microarray analysis, total RNA (100 ng) from each wild-type and mutant embryo was amplified using Two-Cycle Target Labeling (Affymetrix) according to the manufacturerâs instructions. Biotinylated target cRNAs (20 Î¼g) from the 3 wild-type and 3 Tcof1+/- embryos were hybridised to separate GeneChipÂ® Mouse Genome 430 2.0 arrays (Affymetrix), following standard Affymetrix procedures.

ORGANISM(S): Mus musculus

SUBMITTER: Gaye Hattem

PROVIDER: E-GEOD-10167 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Prevention of the neurocristopathy Treacher Collins syndrome through inhibition of p53 function.

Jones Natalie C NC Lynn Megan L ML Gaudenz Karin K Sakai Daisuke D Aoto Kazushi K Rey Jean-Phillipe JP Glynn Earl F EF Ellington Lacey L Du Chunying C Dixon Jill J Dixon Michael J MJ Trainor Paul A PA

Nature medicine 20080203 2

Treacher Collins syndrome (TCS) is a congenital disorder of craniofacial development arising from mutations in TCOF1, which encodes the nucleolar phosphoprotein Treacle. Haploinsufficiency of Tcof1 perturbs mature ribosome biogenesis, resulting in stabilization of p53 and the cyclin G1-mediated cell-cycle arrest that underpins the specificity of neuroepithelial apoptosis and neural crest cell hypoplasia characteristic of TCS. Here we show that inhibition of p53 prevents cyclin G1-driven apoptoti ...[more]

PMID: 18246078

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Project description:Mammalian embryogenesis is characterized by rapid cellular proliferation and diversification. Within a few weeks, a single cell zygote gives rise to millions of cells expressing a panoply of molecular programs, including much of the diversity that will subsequently be present in adult tissues. Although intensively studied, a comprehensive delineation of the major cellular trajectories that comprise mammalian development in vivo remains elusive. For mouse embryogenesis in particular, we and others have performed single cell or single nucleus RNA-seq data (scRNA-seq) during implantation, gastrulation and organogenesis. Here we set out to integrate several single cell RNA-seq datasets (scRNA-seq) that collectively span mouse gastrulation and organogenesis. However, a technical challenge that we faced is that the datasets that we sought to integrate were generated by different groups at different times using different scRNA-seq technologies. In particular, probably because there was no overlapping timepoint, the integration of scRNA-seq data generated at E8.5 (cells, 10X Genomics) and E9.5 (nuclei, sci-RNA-seq3) was challenging (Cao et al. 2019; Pijuan-Sala et al. 2019). To address this, we set out to generate new data at E8.5 that might serve to “bridge” these two datasets. Because of how quickly changes are occurring during this window of development, we focused on individual, somite-resolved E8.5 embryos using a simplified, optimized version of sci-RNA-seq3. We selected 12 embryos from 2 separate litters harvested at E8.5, including a single primitive streak stage embryo (prior to somitogenesis) and 11 embryos staged in 1-somite increments from 2 to 12 somites. The optimized sci-RNA-seq3 method markedly improved data quality, with 9-fold higher UMIs and 6-fold higher gene detection per nucleus, relative to (Cao et al. 2019). Overall, we collected published data (Cheng et al. 2019; Mohammed et al. 2017; Pijuan-Sala et al. 2019), the new E8.5 data, and published data from one study spanning E9.5 to E13.5 but with deeper sequencing of those libraries (Cao et al. 2019). Altogether, we define cell states at each of 19 successive stages spanning E3.5 to E13.5, heuristically connect them to their pseudo-ancestors and pseudo-descendants. Despite being constructed through automated procedures, the resulting trajectories of mammalian embryogenesis (TOME) are largely consistent with our contemporary understanding of mammalian development. In addition, the new E8.5 data itself comprises a foundational resource for mammalian developmental biology (especially for the early somitogenesis), and are made available in a way that will facilitate their ongoing annotation by the research community.

Dataset Information

Transcription profiling of mouse E8.5 wild-type and 3 E8.5 Tcof1+/- littermate embryos (Treacher Collins Syndrome)

Publications

Prevention of the neurocristopathy Treacher Collins syndrome through inhibition of p53 function.

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