RNAseq analysis of A549 cells depleted of RPS19 using siRNAs
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE212775 | GEO | 2022/09/30
REPOSITORIES: GEO
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Project description:We employed next generation sequencing to examine whether knocking down the steroid receptor RNA activator (SRA) gene significantly affect the expression levels of certain genes in MCF-7 cells. MCF-7 cells were transfected with either a pool of four non-target control siRNAs or a pool of four SRA siRNAs for 32 hrs. 157 million reads were generated from triplicate samples of the control group; 151 million reads were generated from triplicate samples of the SRA knockdown group. Six genes were identified as significantly changed in the expression levels with the cutoff of q value ≤ 0.05, fold change ≤ 0.5 or ≥ 2, and reads per kilobase per million mapped reads (RPKM) ≥ 1. However, except for SRA itself, the other five genes were shown by real-time PCR to be only affected by one siRNA in the SRA siRNA pool. Further analysis of this dataset with different cuttoff setting may reveal true SRA-regulated genes in MCF-7.
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Project description:In a previous study, seed coat and cotyledon tissues of Williams, Richland and T157 soybean lines were investigated to show tissue specificity of CHS siRNA expression (Tuteja et al., 2009). Here, we investigated more tissues such as leaf, root and germinating cotyledon to ascertain the tissue specificity of CHS siRNAs in Williams. Data from multiple small RNA libraries were sequenced deeply by the Illumina high-throughput sequencing technology. The total numbers of small RNA reads were from three million to thirty million, providing sufficient data to show the tissue specificity of CHS siRNA. High-throughput sequencing using Genome Analyzer II and Illumina HiSeq 2000 was performed.
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Project description:Purpose: Opium poppy is one of the most important medical plants and remains the only commercial resource of morphinan-based painkillers. However, little is known about its regulation mechanism in benzylisoquinoline alkaloids (BIAs) biosynthesis. Herein, the Transcriptome dataset of Opium poppy was constructed to identify the gene involved in its regulation mechanism in BIAs biosynthesis. Methods: Using Illumina HiSeq X Ten platform, 33 samples of Illumina transcriptome data from different tissues, growth phases and cultivars were constructed. Results: The high-quality transcripts were subsequent quantified with the short reads, and the expression of each unigenes among different samples was calculated by RPKM (the reads per kilobase per million mapped reads). Conclusions: These data provide a foundation of opium poppy transcriptome which may contribute to understand the regulation of BIAs biosynthesis.
2019-04-11 | GSE129566 | GEO
Project description:Purpose: Opium poppy is one of the most important medical plants and remains the only commercial resource of morphinan-based painkillers. However, little is known about its regulation mechanism in benzylisoquinoline alkaloids (BIAs) biosynthesis. Herein, the Transcriptome dataset of Opium poppy was constructed to identify the gene involved in its regulation mechanism in BIAs biosynthesis. Methods: Using Illumina HiSeq X Ten platform, 33 samples of Illumina transcriptome data from different tissues, growth phases and cultivars were constructed. Results: The high-quality transcripts were subsequent quantified with the short reads, and the expression of each unigenes among different samples was calculated by RPKM (the reads per kilobase per million mapped reads). Conclusions: These data provide a foundation of opium poppy transcriptome which may contribute to understand the regulation of BIAs biosynthesis.
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Project description:We employed next generation sequencing to examine whether knocking down the steroid receptor RNA activator (SRA) gene significantly affect the expression levels of certain genes in MCF-7 cells. MCF-7 cells were transfected with either a pool of four non-target control siRNAs or a pool of four SRA siRNAs for 32 hrs. 157 million reads were generated from triplicate samples of the control group; 151 million reads were generated from triplicate samples of the SRA knockdown group. Six genes were identified as significantly changed in the expression levels with the cutoff of q value ≤ 0.05, fold change ≤ 0.5 or ≥ 2, and reads per kilobase per million mapped reads (RPKM) ≥ 1. However, except for SRA itself, the other five genes were shown by real-time PCR to be only affected by one siRNA in the SRA siRNA pool. Further analysis of this dataset with different cuttoff setting may reveal true SRA-regulated genes in MCF-7. MCF-7 cells were cultured in high glucose DMEM with 10% fetal bovine serum, 2 mM Glutamax-1, 100 units/ml penicillin and 100 μg/ml streptomycin. ON-TARGETplus SMARTpool for human SRA (Thermo Scientific, L-027192-00-0005) was used to knockdown SRA (siSRA) and ON-TARGETplus Non-targeting Pool Thermo Scientific, D-001810-10-05) was used as a negative control (siCtrl). A total of 25 nM siRNA was transfected in 6-well dishes using Lipofectamine™ RNAiMAX Reagent (Life Technologies, Invitrogen) following the manufacturer’s recommendations. Polyadenylated RNA was purified from the cells 32 hrs after transfection. cDNA libraries were prepared and double-stranded cDNA was fragmented using DNase I according to Illumina specifications, prior to adaptor ligation. Sequencing libraries were amplified and sequenced using an Illumina HiSeq 2000 sequencer.
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