Whole transcriptome sequencing of mouse cortical astrocytes (DIV 14).
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ABSTRACT: To gain futher insight into the Npas3 regulatory network in astrocytes, RNA-seq was used to analyze the genome-wide changes resulting from the days in vitro (DIV) 14 astrocytes of P0 wild-type (WT) mice and littermate NPAS3-/- mice. Total RNA was extracted from DIV 14 astrocytes of P0 Npas3-/- and WT mice. Then, total RNA was quantified and quality controlled by Agilent Bioanalyzer 2100 system. After cDNA library construction, the library preparations were sequenced on an Illumina NovaSeq 6000 platform in Novogene.
Project description:Purpose: To gain insight into the impact of NPAS3 deficiency on astrogenesis in mice, RNA-seq was used to analyze the genome-wide changes resulting from Npas3+/- and WT cortex at P0 stage. Methods: Total RNA was extracted from Npas3+/- and WT cortex at P0 stage. Then, total RNA was quantified and quality controlled by Agilent Bioanalyzer 2100 system. After cDNA library construction, the library preparations were sequenced on an Illumina NovaSeq 6000 platform in Novogene. Results: At significance cutoffs corresponding to FDR < 0.05, a total of 625 DEGs were identified in Npas3+/- mice. Conclusions: Our results show that Npas3+/- mice exhibit defective astrogenesis and abnormal synapse function.
Project description:To understand the factors influencing gRNA performance for CRISPR-TO system, we performed lncRNA-seq profiling through Novogene Corporation Inc for total RNA extracted from 6 DIV primary mouse cortical neurons dissected from 3 litters of P0 pups of C57BL/6J mice.
Project description:Purpose: To gain futher insight into how STING causes abnormal cortical development in mice ,single cell RNA-seq was used to analyze the cell type changes resulting from the cerebral cortices of P0 STING conditional knock out mice and littermate wild-type. Methods: Single cell isolation was prepared from P0 telencephalic tissue of wild-type(WT) and STINGf/f;Nestin-Cre mice. The fragmented cDNAs containing barcode and UMI sequences was built the library . Agilent 2100 Bioanalyze was used to quality controlled and quantified.Single cell RNA-sequencing analysis was used by the Illumina HiSeq 2500 platform in Annoroad Genomics Results: Clustering and labeling of these cells visualized on a t-SNE plot between the wild-type(WT) and STINGf/f;Nestin-Cre mice brain cortex Conclusions: Single cell RNA-seq data would provide a overall understanding of how STING gene influences cortical development in mice.
Project description:The goal of the study was to compare gene expression of P0 wild-type and P0 Fezf2-/- cortices. Total RNAs were isolated from P0 cortices dissected from wild-type and Fezf2-/- mice (n=3 for each genotype), following Qiagen RNAeasy kit instruction.Sequence libraries were made following Illumina RNA TruSeq library preparation guide.The libaries were pair-end sequenced (50nt per end). Differentially expressed genes were identified by DESEQ.
Project description:The goal of the study was to compare gene expression of P0 wild-type and P0 Satb2-/- cortices. Total RNAs were isolated from P0 cortices dissected from wild-type and Satb2-/- mice (n=3 for each genotype), following Qiagen RNAeasy kit instruction.Sequence libraries were made following Illumina RNA TruSeq library preparation guide.The libaries were pair-end sequenced (50nt per end). Differentially expressed genes were identified by DESEQ.
Project description:Purpose: To gain futher insight into how IFITM2 regulates the neurogenesis ,RNA-seq was used to analyze the genome-wide changes resulting from the cerebral cortices of E13 IFITM2 conditional knock out mice and littermate wild-type. Methods: Total RNA from E13 telencephalic tissue of wild-type(WT) and Ifitm2fl/fl;Nestin-Cre mice was extracted. Specifically, Agilent 2100 Bioanalyze was used to quality controlled and quantified. then, total RNA was converted to cDNA and bound the library. RNA-sequencing analysis was used by the Illumina HiSeq 6000 platform in Annoroad Genomics Approximately one thousand transcripts showed differential expression between the wild-type(WT) and Ifitm2fl/fl;Nestin-Cre mice brain cortex, with a fold change ≥1.5 and p value <0.05. These results reflected IFITM2 plays a role in cortex development. Conclusions: RNA-seq based transcriptome characterization would provide a overall understanding how IFITM2 gene regulated the neurogenesis.
Project description:The experiment was designed to compare transcriptomic differences between WT and Ccr6 KO Tregs during activation. WT and Ccr6 KO Tregs, cells were isolated from mice and cultured in vitro for 3 days with activation using anti-CD3/CD28 beads. Total RNA was extracted using the Trizol method. Quantity and quality were assessed using a Thermo Scientific™ NanoDrop™ 2000/2000c Spectrophotometer. Novogene Corporation Inc prepared the RNA-seq 250-300 bp insert cDNA library. Illumina HiSeq platform PE150 sequencing was used for sequencing, yielding 20M raw reads/sample. Mus Musculus mm39 was used as the reference genome for alignment.