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Next Generation Sequencing Facilitates Quantitative Analysis of Gene expression profile in CD8+ T cells with TOX-overexpression

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of Gene expression profile in CD8+ T cells with TOX-overexpression

PROVIDER: PRJNA492897 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7895510_1.fastq.gz	Fastqsanger.gz
	SRR7895510_2.fastq.gz	Fastqsanger.gz
	SRR7895511_1.fastq.gz	Fastqsanger.gz
	SRR7895511_2.fastq.gz	Fastqsanger.gz
	SRR7895506_1.fastq.gz	Fastqsanger.gz

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Next Generation Sequencing Facilitates Quantitative Analysis of Gene expression profile in CD8+ T cells with TOX-overexpression

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 TOX associated 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. Methods: mRNA profiles of human CD8+ T cell overexpressed by TOX lentivirus or NC lentiviruswere generated by deep sequencing, in triplicate, using Illumina. qRT–PCR validation was performed using TaqMan and SYBR Green assays Conclusions: Our study represents the first detailed analysis of TOX associated transcriptomes 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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