Whole embryo RNA-seq of 73 mouse mutant lines and baseline controls from the Mouse Genetics Project (DMDD)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Illumina HiSeq 2500, Illumina HiSeq 2000
ORGANISM(S): Mus musculus
SUBMITTER: Elisabeth Busch-Nentwich
PROVIDER: E-MTAB-7322 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Publications
Common and distinct transcriptional signatures of mammalian embryonic lethality.
Collins John E JE White Richard J RJ Staudt Nicole N Sealy Ian M IM Packham Ian I Wali Neha N Tudor Catherine C Mazzeo Cecilia C Green Angela A Siragher Emma E Ryder Edward E White Jacqueline K JK Papatheodoru Irene I Tang Amy A Füllgrabe Anja A Billis Konstantinos K Geyer Stefan H SH Weninger Wolfgang J WJ Galli Antonella A Hemberger Myriam M Stemple Derek L DL Robertson Elizabeth E Smith James C JC Mohun Timothy T Adams David J DJ Busch-Nentwich Elisabeth M EM
Nature communications 20190626 1
The Deciphering the Mechanisms of Developmental Disorders programme has analysed the morphological and molecular phenotypes of embryonic and perinatal lethal mouse mutant lines in order to investigate the causes of embryonic lethality. Here we show that individual whole-embryo RNA-seq of 73 mouse mutant lines (>1000 transcriptomes) identifies transcriptional events underlying embryonic lethality and associates previously uncharacterised genes with specific pathways and tissues. For example, our ...[more]
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Project description:2-Dodecyl-6-Methoxycyclohexa-2, 5 -Diene-1,4-Dione (DMDD) was purified from the roots of Averrhoa carambola L. Previous research demonstrated that DMDD is a small molecular compound with significant therapeutic potential for diabetes. However, the efficacy of DMDD to ameliorate diabetic cognitive impairment in type 2 diabetes mice has not been reported. In present study, we detected gene expression profiles in the hippocampi of diabetic db/db mice after DMDD treatment. 9-week-old male db/db mice were treated with DMDD (50 mg/kg) for 28 days followed by Y maze and novel object recognition test for behavioral evaluation. Our results showed that DMDD attenuated the spatial working memory and object recognition memory impairment in db/db mice. Using mouse lncRNA array analysis, 11 lncRNAs and 4 mRNAs with significant differential expression were identified after DMDD treatment. Among these, Hif3a expression was significantly increased in the hippocampi of diabetic db/db mice compared with db/m control mice and then decreased after DMDD treatment. Besides, DMDD treatment reduced tau protein expression, the formation of a-synuclein oligomers and high glucose-induced apoptosis in the hippocampi of db/db mice and high glucose-treated HT22 cells. DMDD protected neural cells from apoptosis by repression of Hif3a. These data suggest that DMDD can alleviate cognitive impairment by regulating apoptosis through the pro-apoptotic protein Hif3a. In conclusion, our study suggests that DMDD has great potential to be a new preventive and therapeutic compound for diabetic cognitive impairment.
2021-12-31 | GSE174378 | GEO
Project description:The DMDD Programme (Deciphering the Mechanisms of Developmental Disorders) provides a free online database of morphological and molecular phenotypes from embryonic-lethal mouse gene knockouts (http://www.dmdd.org.uk/). Embryos are imaged using HREM, placentas are examined by histology and mutant embryo mRNA expression profiles are compared to wild type. Total RNA was extracted from somite number staged homozygous mutant, heterozygous and sibling wild-type E9.5 whole C57BL/6N embryos and DNase treated. The embryos were derived from Mouse Genetics Programme gene knockout lines defined as homozygous embryonic lethal (http://www.sanger.ac.uk/science/collaboration/mouse-resource-portal). Stranded RNA-seq libraries were constructed using the Illumina TruSeq Stranded RNA protocol with oligo dT pulldown.<br> Notes about samples and libraries: </br><br>(1) The knockout alleles in this project were: Cnot4: Cnot4^tm1b(EUCOMM)Wtsi ; Ssr2: Ssr2^tm1b(EUCOMM)Wtsi ; 4933434E20Rik: 4933434E20Rik^tm1a(EUCOMM)Wtsi ; 1700007K13Rik: 1700007K13Rik^tm2b(EUCOMM)Wtsi ; Camsap3: Camsap3^tm1a(EUCOMM)Wtsi ; Anks6: Anks6^tm1b(KOMP)Wtsi ; Pdzk1: Pdzk1^tm2b(EUCOMM)Wtsi ; Otud7b: Otud7b^tm1b(EUCOMM)Wtsi ; Adamts3: Adamts3^tm1b(KOMP)Wtsi; Mir96: Mir96^tm2.2WTSI ; Cyp11a1: Cyp11a1^tm1b(EUCOMM)Hmgu and Traf6: Traf6^tm2a(EUCOMM)Wtsi. </br><br> (2) A combination of litter identifier and embryo identifier within a litter will unambiguously identify a single embryo used in this study. </br><br> (3) There is a margin of error for the somite-stage information (+/- 1 somite) because some embryos could be in between somite stages.</br><br> (4) There are a minimum of three biological replicates per line. </br><br> (5) Sex of the embryos was determined post-RNA-seq by looking at the expression of Xist (strong expression in females only). </br><br> (6) Libraries were constructed in batches comprising multiple litters from one line from RNA-extraction to sequencing.</br><br> This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ .</br>
2015-02-11 | E-MTAB-5224 | biostudies-arrayexpress
Project description:The DMDD Programme (Deciphering the Mechanisms of Developmental Disorders ) provides a free online database of morphological and molecular phenotypes from embryonic-lethal mouse gene knockouts (http://www.dmdd.org.uk/). Embryos are imaged using high-resolution episcopic microscopy (HREM), placentas are examined by histology and mutant embryo mRNA expression profiles are compared to wild type. To underpin these investigations we have produced a comprehensive time series of gene expression through normal embryo development<br></br>In this study, yotal RNA was extracted from wildtype C57BL/6N sibling embryos with heterozygous parentage from the Mouse Genetics Project (http://www.dmdd.org.uk/) and DNase treated. Stranded RNAseq libraries were constructed using the Illumina TruSeq Stranded RNA protocol with oligo dT pulldown. <br></br> Notes about samples and libraries:<br></br> (1) A combination of litter identifier and embryo identifier within a litter will unambiguously identify a single embryo used in this study. <br></br> (2) There is a margin of error for the somite-stage information (+/- 1 somite) because some embryos could be in between somite stages. <br></br>(3) Sex of the embryos was determined post-RNA-seq by looking at the expression of Xist (strong expression in females only). <br></br>(4) All knockouts in the parents (and hence grandparents) are for embryonic lethal genes. <br></br> (5) There are four biological replicates per somite-stage. <br></br>(6) Complementary data on genotypically wild-type mice with mixed G0 lineage and no history of genetic modification can also be found in ArrayExpress under accession numbers E-ERAD-401 ( https://www.ebi.ac.uk/arrayexpress/experiments/E-ERAD-401 ) and E-ERAD-499 ( https://www.ebi.ac.uk/arrayexpress/experiments/E-ERAD-499 ) respectively.
2015-02-11 | E-ERAD-352 | biostudies-arrayexpress
Project description:The DMDD Programme (Deciphering the Mechanisms of Developmental Disorders, https://dmdd.org.uk/) provides a free online database of morphological and molecular phenotypes from embryonic-lethal mouse gene knockouts (http://www.dmdd.org.uk/). Embryos are imaged using HREM, placentas are examined by histology and mutant embryo mRNA expression profiles are compared to wild type. To underpin these investigations we have produced a comprehensive time series of gene expression through normal embryo development. Total RNA was extracted from somite number staged, second generation genotypically wild type, C57BL/6N embryos of mixed G0 linages from the Mouse Genetics Programme (http://www.sanger.ac.uk/science/collaboration/mouse-resource-portal) and DNase treated. Stranded RNA-seq libraries were constructed using the Illumina TruSeq Stranded RNA protocol with oligo dT pulldown.<br> Notes about samples and libraries: </br><br>(1) A combination of litter identifier and embryo identifier within a litter will unambiguously identify a single embryo used in this study. </br><br>(2) There is a margin of error for the somite-stage information (+/- 1 somite) because some embryos could be in between somite stages. </br><br> (3) All knockouts in the parents or grandparents are for embryonic lethal genes. </br><br>(4) There are four biological replicates per somite-stage, with the exception of the 4-somite embryos (three biological replicates only). Altogether 111 embryos were sourced, of which 14 were sequenced twice to generate enough read depth/coverage. No new libraries were prepared for the repeated sequencing, despite the assays being assigned new ERX* accessions. </br><br> (5) Sex of the embryos was determined post-RNA-seq by looking at the expression of Xist (strong expression in females only). </br><br> (6) Library constructon batch refers to batch of sample handling post RNA-extraction (size selection, PCR amplification during library preparataion). </br><br>(7) Superbatch gathers 17 representative embryos out of the 111 embryos post RNA-extraction, and put them through the library construction pipeline as one single batch. The generated data is intended to be used for normalisation using the remove unwanted variation (RUV) method. </br> <br> This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ .</br>
2015-11-03 | E-ERAD-401 | biostudies-arrayexpress
Project description:Objective DLL1 is a mammalian Notch ligand with essential functions during embryonic development. We tagged endogenous DLL1 in one homologous recombination step such that AcGFP-HA tagged DLL1 could be converted by Cre-mediated site-specific recombination into StrepFlag tagged DLL1. We anticipated that this should allow us to visualise DLL1 in living cells as well as allow for sorting and enrichment of DLL1-expressing cells and efficient purification of DLL1 protein complex. Results We generated constructs to express a DLL1 variant that carried C-terminal in frame an AcGFPHA tag flanked by loxP sites followed by a StrepFlag tag out of frame. Cre-mediated recombination removed AcGFP-HA and added StrepFlag in frame to DLL1. The AcGFPHAstopStrepFlag tag was added to the Dll1 cDNA to allow for tests in cultured cells in vitro and was introduced into endogenous DLL1 in mice using ES cells modified by homologous recombination. Tagged DLL1 protein was detected by antibodies against GFP and HA or Flag, respectively, both in CHO cell and embryo lysates. In CHO cells the AcGFP protein fused to DLL1 was functional. In vivo AcGFP expression was below the level of detection by direct fluorescence. However, the StrepFlag tag allowed us to specifically purify DLL1 complexes from embryo lysates. Homozygous mice expressing AcGFPHA or StrepFlag-tagged DLL1 revealed a vertebral column phenotype reminiscent of disturbances in AP polarity during somitogenesis, a process most sensitive to reduced DLL1 function. Thus, even small C-terminal tags can impinge on sensitive developmental processes requiring DLL1 activity.
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Project description:The DMDD Programme (Deciphering the Mechanisms of Developmental Disorders ) provides a free online database of morphological and molecular phenotypes from embryonic-lethal mouse gene knockouts (http://www.dmdd.org.uk/). Embryos are imaged using HREM, placentas are examined by histology and mutant embryo mRNA expression profiles are compared to wild type. To underpin these investigations we have produced a comprehensive time series of gene expression through normal embryo development. Total RNA was extracted from somite number staged wildtype embryos with no history of genetic modification in previous generations from the Mouse Genetics Programme (http://www.sanger.ac.uk/science/collaboration/mouse-resource-portal) and DNase treated. Stranded RNA-seq libraries were constructed using the Illumina TruSeq Stranded RNA protocol with oligo dT pulldown.<br> Notes about samples and libraries:<br> (1) A combination of litter identifier and embryo identifier within a litter will unambiguously identify a single embryo used in this study. </br><br> (2) There is a margin of error for the somite-stage information (+/- 1 somite) because some embryos could be in between somite stages. </br><br> (3) Sex of the embryos was determined post-RNA-seq by looking at the expression of Xist (strong expression in females only). </br><br> (4) Complementary data on genotypically wild-type mice with mixed G0 lineage and with heterozygous mutant parents can also be found in ArrayExpress under accession numbers E-ERAD-352 ( https://www.ebi.ac.uk/arrayexpress/experiments/E-ERAD-352 ) and E-ERAD-401 ( https://www.ebi.ac.uk/arrayexpress/experiments/E-ERAD-401 ).</br>
2016-06-01 | E-ERAD-499 | biostudies-arrayexpress
Project description:Pathogenic variants in ACTN2, coding for alpha-actinin 2, are known to be rare causes of Hyper-trophic Cardiomyopathy. However, little is known about the underlying disease mechanisms. Adult heterozygous mice carrying the Actn2 M228T variant were phenotyped by echocardiog-raphy. For homozygous mice, viable E15.5 embryonic hearts were analysed by High Resolution Episcopic Microscopy and wholemount staining, complemented by unbiased proteomics, qPCR and Western blotting. Heterozygous Actn2 M228T mice have no overt phenotype. Only mature males show molecular parameters indicative of cardiomyopathy. By contrast, the variant is em-bryonically lethal in the homozygous setting and E15.5 hearts show multiple morphological ab-normalities. Molecular analyses, including unbiased proteomics, identified quantitative abnormal-ities in sarcomeric parameters, cell cycle defects and mitochondrial dysfunction. The mutant al-pha-actinin protein is found to be destabilised, associated with increased activity of the ubiqui-tin-proteosomal system. This missense variant in alpha-actinin renders the protein less stable. In response, the ubiquitin-proteosomal system is activated; a mechanism which has been implicated in cardiomyopathies previously. In parallel, lack of functional alpha-actinin is thought to cause energetic defects through mitochondrial dysfunction. This seems, together with cell cycle defects, the likely cause of death of the embryos. The defects also have wide-ranging morphological con-sequences.
2023-05-10 | PXD039226 | Pride
Project description:drl expression initiates during gastrulation and condenses as a band of cells at the prospective lateral embryo margin. In late epiboly, drl:EGFP is detectable as a band of scattered EGFP-fluorescent cells; after gastrulation, drl:EGFP-positive cells coalesce at the embryo margin that then in somitogenesis break down into the anterior and posterior lateral plate with subsequent cell migrations that form the posterior vascular/hematopoietic stripes and the anterior cardiovascular and myeloid precursors. Data is as published in Mosimann & Panakova et al (Nature Communications, 2015) The same pool of homozygous drl:EGFP transgenic zebrafish were mass-mated 3 times and embryos collected at 2ss. Embryos were de-chorionated, homogenized, and sorted, then subject to mRNA isolation and expression analysis.
2015-07-14 | E-GEOD-70881 | biostudies-arrayexpress
Project description:LYVE-1-positive macrophages were observed to be closely spatially associated with the developing lymphatic vasculature. The role of this population of macrophages in the embryo is uncharacterised. We used microarray analyses to investigate which genes are differentially regulated between LYVE-1-positive and LYVE-1-negative macrophages Macrophage populations were isolated from cell suspensions prepared from digested embryonic day (E) 15.5 skin and prepared for RNA extraction and hybridization on Affymetrix microarrays.
2010-10-02 | E-GEOD-24492 | biostudies-arrayexpress