Abstract
A detailed understanding of post-transcriptional gene expression is necessary to correlate the different elements involved in the many levels of RNA-protein interactions that are needed to coordinate the cellular biomolecular machinery. The profile of mRNA, a major component of this machinery, can be examined after isolation from specific RNA-binding proteins (RBPs). RIP-Chip or ribonomic profiling is a versatilein vivo technique that has been widely used to study post-transcriptional gene regulation and the localization of mRNA. Here we elaborately detail the methodology for mRNA isolation using RBP immunoprecipitation (RIP) as a primary approach. Specific antibodies are used to target RBPs, which are then used to capture the associated mRNA.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Animals
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Antibodies, Immobilized / immunology
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Antibodies, Immobilized / metabolism
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Antibody Specificity
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Cell Extracts
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Computational Biology
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HeLa Cells
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Humans
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Immunoprecipitation / methods*
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K562 Cells
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Magnetics
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Microspheres
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Nerve Tissue Proteins / metabolism
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Oligonucleotide Array Sequence Analysis
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RNA Transport
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RNA, Messenger / analysis*
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RNA, Messenger / isolation & purification
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RNA, Messenger / metabolism
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Ribonucleoproteins / immunology
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Ribonucleoproteins / isolation & purification
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Ribonucleoproteins / metabolism
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Staphylococcal Protein A / metabolism
Substances
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Antibodies, Immobilized
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Cell Extracts
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G-substrate
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Nerve Tissue Proteins
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RNA, Messenger
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Ribonucleoproteins
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Staphylococcal Protein A