Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq of E10.5 limb mesenchymal cells from mice with mutations in Mycn and mir17-92

ABSTRACT: Mice with conditionally deactivating mutations in Mycn (Mycnfl/fl) or mir17-92 & mir106b (mir17-92fl/fl ; mir106b-/-) were crossed with mice expressing Cre-recombinase driven by the Prx1 promoter (Prx1Cre). Embryos were collected at E10.5, and distal compartment of hind- and fore-limbs were isolated from wild-type and conditional knock-out embryos (n=3 replicates per biologic group). Total RNA was extracted from each specimen with Trizol, and used in RNA-seq library preparation with Illumina Truseq mRNA library kit. Libraries were sequenced on the Illumina NextSeq system with 75bp single end reads.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Mus musculus

SUBMITTER: Ugur Ayturk

PROVIDER: E-MTAB-5671 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Detailed information about stage-specific changes in gene expression is crucial for understanding the gene regulatory networks underlying development and the various signal transduction pathways contributing to morphogenesis. Here, we describe the global gene expression dynamics during early murine limb development, when cartilage, tendons, muscle, joints, vasculature, and nerves are specified and the musculoskeletal system of the limbs is established. We used whole-genome microarrays to identify genes with differential expression at 5 stages of limb development (E9.5 to 13.5), during fore-limb and hind-limb patterning. We found that the onset of limb formation is characterized by an up-regulation of transcription factors, which is followed by a massive activation of genes during E10.5 and E11.5 which tampers off at later time points. Among 3520 genes identified as significantly up-regulated in the limb, we find ~30% to be novel, dramatically expanding the repertoire of candidate genes likely to function in the limb. Hierarchical and stage-specific clustering identified expression profiles that correlate with functional programs during limb development and are likely to provide new insights into specific tissue patterning processes. Here we provide for the first time, a comprehensve analysis of developmentally regulated genes during murine limb development, and provide some novel insights into the expression dynamics governing limb morphogenesis. Fifty- one arrays were analyzed, consisting of whole fore-limb and hind-limb bud RNA (experimental) and whole embryo RNA (reference) samples from E9.5 to E13.5 DPC mouse (FVB strain). Embryos were not pooled to generate samples. Each time point has 3 to 5 biological replicates for limb bud samples, duplicates for whole embryos. Comparisons were made between limb bud samples and whole embryo at the same stage, fore-limb samples of different stages, hind-limb samples of different stages, and fore-limb samples compared to hind-limb samples at the same or the next stage.

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RNA-seq of E10.5 limb mesenchymal cells from mice with mutations in Mycn and mir17-92

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