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Ribosome profiling : ウィキペディア英語版 | Ribosome profiling
Ribosome profiling, or Ribo-Seq, is a technique developed by Nick Ingolia and Jonathan Weissman that uses specialized messenger RNA (mRNA) sequencing to determine which mRNAs are being actively translated. It produces a “global snapshot” of all the ribosomes active in a cell at a particular moment, called as translatome. Consequently, this enables researchers to identify the location of translation start sites, their distribution, and the speed of the translating ribosomes. Ribosome profiling involves similar sequencing library preparation and data analysis to RNA-Seq, but unlike RNA-Seq, which sequences all of the mRNA of a given sequence present in a sample, ribosome profiling targets only mRNA sequences protected by the ribosome during the process of decoding by translation.〔 ==History== Ribosome profiling is based on the discovery that the mRNA within a ribosome can be isolated through the use of nucleases that degrade unprotected mRNA regions. This technique analyzes the regions of mRNAs being converted to protein, as well as the levels of translation of each region to provide insight into global gene expression. Prior to its development, efforts to measure translation in vivo included microarray analysis on the RNA isolated from polysomes, as well as translational profiling through the affinity purification of epitope tagged ribosomes. These are useful and complementary methods, but neither allows the sensitivity and positional information provided by ribosome profiling.〔
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