Next Generation Sequencing Facilitates Quantitative Analysis of oral epithelial cells from SPF 8 weeks and 1 week old Wild Type mice
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE161973 | GEO | 2021/01/21
REPOSITORIES: GEO
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Project description:Purpose: RNAseq analysis to identify differentiatlly expressed genes due to the presnce of microbiota
2021-01-21 | GSE161974 | GEO
Project description:Next Generation Sequencing Facilitates Quantitative Analysis of oral epithelial cells from SPF 8 weeks and 1 week old Wild Type mice
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Project description:Unlike adult β-cells, fetal and neonatal islets are functional immature and have blunted glucose responsiveness and decreased insulin secretion in response to stimuli. Pancreatic islets are a mixture of different cell types. Study of transcriptomes from intact islets would identify novel molecular mechanisms controlling islet functional development. At e19 and 2 weeks of age, pancreatic islets were isolated and total RNA were extracted for RNA-Seq study. Gene expression profiles were compared in the study.
2020-07-02 | GSE153604 | GEO
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived femoral diaphysis and metaphysis transcriptome profiling (RNA-seq) to determine pathways and networks dependent on Dlx3 during bone development and homeostasis. Methods: mRNA profiles of diaphysis and metaphysis isolated from the femur of 5-week-old wild-type (WT) and Dlx3Oc-cKO (OC-cre;Dlx3f/-) conditional knockout mice were generated by deep sequencing, in triplicate, using Illumina HiSeq 2000. The sequence reads that passed quality filters were analyzed at the transcript isoform level by ANOVA (ANOVA) and TopHat. qRT-PCR validation was performed using SYBR Green assay. Results: RNA-Seq data were generated with Illumina's HiSeq 2000 system. Raw sequencing data were processed with CASAVA 1.8.2 to generate fastq files. Reads of 50 bases were mapped to the mouse transcriptome and genome mm9 using TopHat 1.3.2. Gene expression values (RPKM) were calculated with Partek Genomics Suite 6.6, which was also used for the ANOVA analysis to determine significantly differentially expressed genes. Conclusions: Our study represents the first detailed analysis of Dlx3Oc-cKO diaphysis and metaphysis from femurs, with biologic triplicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions. Diaphysis and metaphysis mRNA profiles of metaphysis and diaphysis from femurs of 5-wk-old (WT) and Dlx3Oc-cKO male mice were generated by deep sequencing, in triplicate, using Illumina HiSeq 2000.
2014-11-04 | E-GEOD-53105 | biostudies-arrayexpress
Project description:Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived normal human kidney transcriptome profiling (RNA-seq) to microarray and quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis The kidney tissue was immediately placed and stored in RNAlater® (Ambion), according to the manufacturer’s instruction. The tissue was manually microdissected under microscope in RNAlater® pool for glomerular and tubular compartment. Dissected tissue was homogenized and RNA was prepared using RNAeasy mini columns (Qiagen, Valencia, CA, US), according to the manufacturer’s instructions. RNA quality and quantity were determined using the Laboratory-on-Chip Total RNA PicoKit Agilent BioAnalyzer. Only samples without evidence of degradation were further used (RNA Integrity Number >6).
2014-08-06 | E-GEOD-60119 | biostudies-arrayexpress
Project description:PurposeNext-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived retinal transcriptome profiling (RNA-seq) to microarray and quantitative reverse transcription polymerase chain reaction (qRT-PCR) methods and to evaluate protocols for optimal high-throughput data analysis.MethodsRetinal mRNA profiles of 21-day-old wild-type (WT) and neural retina leucine zipper knockout (Nrl(-/-)) mice were generated by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows-Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT-PCR validation was performed using TaqMan and SYBR Green assays.ResultsUsing an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to the mouse genome (build mm9) and identified 16,014 transcripts in the retinas of WT and Nrl(-/-) mice with BWA workflow and 34,115 transcripts with TopHat workflow. RNA-seq data confirmed stable expression of 25 known housekeeping genes, and 12 of these were validated with qRT-PCR. RNA-seq data had a linear relationship with qRT-PCR for more than four orders of magnitude and a goodness of fit (R(2)) of 0.8798. Approximately 10% of the transcripts showed differential expression between the WT and Nrl(-/-) retina, with a fold change ≥1.5 and p value <0.05. Altered expression of 25 genes was confirmed with qRT-PCR, demonstrating the high degree of sensitivity of the RNA-seq method. Hierarchical clustering of differentially expressed genes uncovered several as yet uncharacterized genes that may contribute to retinal function. Data analysis with BWA and TopHat workflows revealed a significant overlap yet provided complementary insights in transcriptome profiling.ConclusionsOur study represents the first detailed analysis of retinal transcriptomes, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a cell or tissue. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.
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Project description:H5N1 highly pathogenic avian influenza (HPAI) has raised global concern for causing huge economic losses in poultry industry. By using reversal genetic technical, we have rescued a H5 wild-type virus rS and its NS1-truncated virus S-NS99, harboring only 99 amino acids in the amino terminus of NS1. We then used a high-throughput RNA-seq method to analyze host and pathogen transcriptomes in the lungs of chickens infected with rS and S-NS99. Sequenced numbers of viral transcripts and expression levels of host immune-related genes at 1 day post infection (dpi) were higher in rS-infected than S-NS99-infected chickens. The numbers of immune response of rS-infected chickens by Gene Ontology (GO) enrichment were higher than S-NS99-infected chickens. KEGG pathway analysis revealed rS infection could provoke significantly stronger immune response than S-NS99. Collectively, our data provide new insight into the underlying mechanisms of the differential length NS1 of avian influenza viruses.
2018-05-25 | GSE114880 | GEO
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare Young and Old muscles transcriptome profiling (RNA-seq) to optimal high-throughput data analysis Results: Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to the mouse genome (build mm10) and identified 24799 genes. Among them, 95.7% of the genes displayed the similar expression level in both young and old muscles.2% of genes (501) were significantly up-regulated and 2.3% of genes (584) were significantly down-regulated. p value <0.05.
2018-03-07 | GSE78147 | GEO
Project description:We used RNAseq to analyse the transcriptomes of cycling and quiescent livers from cTKO (Rblox/lox;p130lox/lox;p107?/?) and control mice. mRNA profiles of cycling and quiescent control and TKO liver tissue were generated by deep sequencing in duplicate
2014-07-12 | E-GEOD-43631 | biostudies-arrayexpress
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular functions. The goal of this study is to compare NGS-derived salivary gland transcriptome profilings (RNA-seq) to better understand the molecular changes in adult sublingual glands in the absence of the Nkx2.3 transcription factors. Methods: Female sublingual salivary gland mRNA profiles were generated by deep sequencing, in 4 replicates for Nkx2.3 knockout mice, using Illumina. The sequence reads that passed quality filters were analyzed at the gene level with STAR followed by DESeq2. Results: Using an optimized data analysis workflow, we mapped about 20 million sequence reads per sample to the mouse genome (build mm9) and identified 496 genes that were differentially expressed between the wildtype and Nkx2.3-knockout murine female sublingual salivary glands . RNA-seq data provided valuable insights into the nature of the functional differences resulting from the Nkx2.3 disruption Conclusions: Our study represents the first detailed analysis of sublingal salivary gland transcriptome profiling differences resulting from the Nkx2.3 disruption, with biologic replicates, generated by RNA-seq technology. Our results confirmed functions of many previously studied genes. We conclude that RNA-seq-based transcriptome characterization would offer a comprehensive and sensitive evaluation of the gene expression.
2023-01-31 | GSE168253 | GEO