Reference: Watson PY and Fedor MJ (2009) Determination of intracellular RNA folding rates using self-cleaving RNAs. Methods Enzymol 468:259-86

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Abstract


We have developed a system that relies on RNA self-cleavage to report quantitatively on assembly of RNA structures in vivo. Self-cleaving RNA sequences are inserted into mRNAs or snoRNAs and expressed in yeast under the control of a regulated promoter. Chimeric RNAs that contain self-cleaving ribozymes turn over faster than chimeric RNAs that contain a mutationally inactivated ribozyme by an amount that reflects the rate at which the ribozyme folds and self-cleaves. A key feature of this system is the choice of assay conditions that selectively monitor intracellular assembly and self-cleavage by suppressing further ribozyme activity during the analysis.

Reference Type
Journal Article | Research Support, N.I.H., Extramural
Authors
Watson PY, Fedor MJ
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