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RNA Sequencing Facilitates Quantitative Analysis of Wild Type and HRD1-/- liver Transcriptomes

ABSTRACT: RNA Sequencing Facilitates Quantitative Analysis of Wild Type and HRD1-/- liver Transcriptomes

PROVIDER: PRJNA450315 | ENA |

REPOSITORIES: ENA

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RNA Sequencing Facilitates Quantitative Analysis of Wild Type and HRD1-/- liver Transcriptomes

Project description:Screen the differential genes of Wild Type and HRD1-/- liver under fasted and refed condition

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Next Generation Sequencing Facilitates Quantitative Analysis of PIGS-KO, PIGS-HRD1-DKO, PIGS-HRD1-CD55-TKO, and PIGS-HRD1-CD55-TKO+HA-CD55 Transcriptomes

Project description:Next Generation Sequencing Facilitates Quantitative Analysis of PIGS-KO, PIGS-HRD1-DKO, PIGS-HRD1-CD55-TKO, and PIGS-HRD1-CD55-TKO+HA-CD55 Transcriptomes

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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and Hrd1-/- T regulatory cells

Project description:Purpose: Natural T regulatory cells (nTreg) and induced Tregs (iTreg) were isolated from hrd1 WT and knockout mice. The purpose of this project is to look at the transcriptional profiles changes of genes specifically involved in T cell receptor signaling, TGF-beta signaling, and metabolic genes in Hrd1 deficient Tregs. Methods: natural Tregs were sorted according to CD4+GFP+, and RNA was harvested using QIAGEN Rneasy Mini Kit. For induced iTreg, CD4 T cells were isolated and polarized with IL2/TGFbeta for 5 days to induce FoxP3 induction. Day 5, GFP positive cells were isolated and RNA were harvested using QIAGEN RNeasy mini Kit. Results and Conclusions: Our study represents the first detailed analysis of transcriptional profiles in Hrd1 deficient T regulatory cells, 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.

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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and Hrd1-/- T regulatory cells

Project description:Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and Hrd1-/- T regulatory cells

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Next Generation Sequencing Facilitates Quantitative Analysis of PIGS-KO, PIGS-HRD1-DKO, PIGS-HRD1-CD55-TKO, and PIGS-HRD1-CD55-TKO+HA-CD55 Transcriptomes

Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare the genes expression difference at transcriptome level; Methods: Total RNA was extracted from whole cells with the mirVana miRNA Isolation Kit according to the manufacturer’s protocol. RNA quality and integrity were evaluated with an Agilent 2100 Bioanalyzer. Samples with an RNA integrity number (RIN) ≥ 7 were considered to be of high quality and were processed further and subjected to subsequent analysis. Total RNA-seq libraries were generated using 4 μg of total RNA, which was analyzed using the TruSeq Stranded mRNA LTSample Prep Kit. These libraries were then sequenced using the Illumina sequencing platform (HiSeqTM 2500 or Illumina HiSeq X Ten), and 125-bp/150-bp paired-end reads were generated. Results: The raw reads containing adaptors and the low-quality reads from the raw data were removed using Trimmomatic to obtain clean reads. Transcriptome sequencing was conducted by OE Biotech Co., Ltd. (Shanghai, China), and clean reads were provided. The clean reads were mapped to the hg38 reference genome using hisat2 (version 2.1.0). The output BAM files were converted to SAM files using SAMtools 1.9. The final TPM values were obtained using Stringtie 1.3.5. Conclusions: To understand the mechanistic basis of GPI biosynthesis upregulation by the CD55 precursor, we performed RNA- sequencing (RNA-seq) of samples of parental PIGS-HRD1-DKO, PIGS-HRD1-CD55-TKO, and PIGS-HRD1-CD55-TKO stably overexpressed HA-CD55 stably overexpressing cells. Total RNA was extracted and analyzed. The expression profile of GPI biosynthesis -related genes was not significantly affected by CD55.

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Next Generation Sequencing Facilitates Quantitative Analysis of Solanum lycopersicum bud and flower Transcriptomes

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RNA Sequencing Facilitates Quantitative Analysis of LINC00901 overexpression and Control MIA PaCa-2 Cell Transcriptomes

Project description:RNA Sequencing Facilitates Quantitative Analysis of LINC00901 overexpression and Control MIA PaCa-2 Cell Transcriptomes

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