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RNA-Seq
RNA-seq (RNA sequencing), also called whole transcriptome shotgun sequencing (WTSS), is a technology that uses the capabilities of next-generation sequencing to reveal a snapshot of RNA presence and quantity from a genome at a given moment in time.
==Introduction==
The transcriptome of a cell is dynamic; it continually changes. The recent developments of next-generation sequencing (NGS) allow for increased base coverage of a DNA sequence, as well as higher sample throughput. This facilitates sequencing of the RNA transcripts in a cell, providing the ability to look at alternative gene spliced transcripts, post-transcriptional modifications, gene fusion, mutations/SNPs and changes in gene expression. In addition to mRNA transcripts, RNA-Seq can look at different populations of RNA to include total RNA, small RNA, such as miRNA, tRNA, and ribosomal profiling. RNA-Seq can also be used to determine exon/intron boundaries and verify or amend previously annotated 5’ and 3’ gene boundaries. Ongoing RNA-Seq research includes observing cellular pathway alterations during infection, and gene expression level changes in cancer studies. Prior to NGS, transcriptomics and gene expression studies were done with expression microarrays, which contain thousands of DNA sequences (probes) that potentially match complementary sequences in the sample, making available a profile of all transcripts being expressed. This was later done with serial analysis of gene expression (SAGE).
Microarrays rely on a good knowledge of an organism's genome. One deficiency with microarrays that makes RNA-Seq more attractive has been limited coverage; such arrays target the identification of known common alleles that represent approximately 500,000 to 2,000,000 SNPs of the more than 10,000,000 in the genome.〔(【引用サイトリンク】title=HapMap: About the Project )〕 As a result, libraries aren’t usually available to detect and evaluate rare allele variant transcripts, and the arrays are only as good as the SNP databases they’re designed from, so they have limited application for research purposes. Many cancers for example are caused by rare <1% mutations and would go undetected. However, arrays still have a place for targeted identification of already known common allele variants, making them ideal for regulatory body-approved diagnostics such as cystic fibrosis.
抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』
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