Isolation of RNP granules
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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Isolation of RNP granules. / Jønson, Lars; Nielsen, Finn Cilius; Christiansen, Jan.
RNA: methods and protocols. ed. / Henrik Nielsen. Vol. 703 Springer, 2011. p. 265-273 (Methods in Molecular Biology).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Isolation of RNP granules
AU - Jønson, Lars
AU - Nielsen, Finn Cilius
AU - Christiansen, Jan
PY - 2011
Y1 - 2011
N2 - The post-transcriptional operon provides a means of synexpression of mRNAs encoding interrelated proteins. The coordination of gene expression may be achieved by a trans-acting RNA-binding protein attaching to similar cis-elements in different, yet functionally clustered, mRNAs. The RNP granule can be regarded as a supramolecular assembly of RNA and protein, probably representing several overlapping post-transcriptional operons. The present protocol describes how RNP granules may be isolated by the transgenic expression of a 3X FLAG version of an RNA-binding protein under tetracycline control via the tetracycline receptor/operator complex. In this way, inclusion of an appropriate tetracycline concentration ensures expression of the tagged version at the endogenous level, and the 3X FLAG tag is a convenient "handle" for the subsequent immunoprecipitation by immobilized anti-FLAG antibody.
AB - The post-transcriptional operon provides a means of synexpression of mRNAs encoding interrelated proteins. The coordination of gene expression may be achieved by a trans-acting RNA-binding protein attaching to similar cis-elements in different, yet functionally clustered, mRNAs. The RNP granule can be regarded as a supramolecular assembly of RNA and protein, probably representing several overlapping post-transcriptional operons. The present protocol describes how RNP granules may be isolated by the transgenic expression of a 3X FLAG version of an RNA-binding protein under tetracycline control via the tetracycline receptor/operator complex. In this way, inclusion of an appropriate tetracycline concentration ensures expression of the tagged version at the endogenous level, and the 3X FLAG tag is a convenient "handle" for the subsequent immunoprecipitation by immobilized anti-FLAG antibody.
U2 - 10.1007/978-1-59745-248-9_18
DO - 10.1007/978-1-59745-248-9_18
M3 - Book chapter
SN - 978-1-58829-913-0
VL - 703
T3 - Methods in Molecular Biology
SP - 265
EP - 273
BT - RNA
A2 - Nielsen, Henrik
PB - Springer
ER -
ID: 40966820