RNA immunoprecipitation for determining RNA-protein associations in vivo.
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RNA immunoprecipitation for determining RNA-protein associations in vivo. / Gilbert, Chris; Svejstrup, Jesper Q.
In: Current Protocols in Molecular Biology, Vol. 75, No. 1, Chapter 27, 08.2006.Research output: Contribution to journal › Review › Research › peer-review
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TY - JOUR
T1 - RNA immunoprecipitation for determining RNA-protein associations in vivo.
AU - Gilbert, Chris
AU - Svejstrup, Jesper Q.
PY - 2006/8
Y1 - 2006/8
N2 - Similar to chromatin immunoprecipitation (ChIP), RNA immunoprecipitation (RIP) can be used to detect the association of individual proteins with specific nucleic acid regions, in this case on RNA. Live cells are treated with formaldehyde to generate protein-RNA cross-links between molecules that are in close proximity in vivo. RNA sequences that cross-link with a given protein are isolated by immunoprecipitation of the protein, and reversal of the formaldehyde cross-linking permits recovery and quantitative analysis of the immunoprecipitated RNA by reverse transcription PCR. The basics of RIP are very similar to those of ChIP, but with some important caveats. This unit describes the RIP procedure for Saccharomyces cerevisiae. Although the corresponding steps for metazoan cells have not yet been worked out, it is likely that the yeast procedure can easily be adapted for use in other organisms.
AB - Similar to chromatin immunoprecipitation (ChIP), RNA immunoprecipitation (RIP) can be used to detect the association of individual proteins with specific nucleic acid regions, in this case on RNA. Live cells are treated with formaldehyde to generate protein-RNA cross-links between molecules that are in close proximity in vivo. RNA sequences that cross-link with a given protein are isolated by immunoprecipitation of the protein, and reversal of the formaldehyde cross-linking permits recovery and quantitative analysis of the immunoprecipitated RNA by reverse transcription PCR. The basics of RIP are very similar to those of ChIP, but with some important caveats. This unit describes the RIP procedure for Saccharomyces cerevisiae. Although the corresponding steps for metazoan cells have not yet been worked out, it is likely that the yeast procedure can easily be adapted for use in other organisms.
U2 - 10.1002/0471142727.mb2704s75
DO - 10.1002/0471142727.mb2704s75
M3 - Review
C2 - 18265380
AN - SCOPUS:39849085276
VL - 75
JO - Current Protocols in Molecular Biology
JF - Current Protocols in Molecular Biology
SN - 1934-3639
IS - 1
M1 - Chapter 27
ER -
ID: 331031080