Abstract
RNA processing by the splicing machinery removes intronic sequences from pre-mRNA to generate mature mRNA transcripts. Many splicing events occur co-transcriptionally when the pre-mRNA is still associated with the transcription machinery. This mechanism raises questions regarding the number of spliceosomes associated with the pre-mRNA at a given time. In this protocol, we present a quantitative FISH approach that measures the ratio of intensities between two different spliceosomal components associated on a nascent mRNA, and compares to the number of introns in the mRNA, thereby calculating the number of spliceosome complexes assembled with each transcript.
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Acknowledgement
The work in the Shav-Tal lab is supported by the European Research Council (ERC).
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Neufeld, N., Brody, Y., Shav-Tal, Y. (2014). Quantifying the Ratio of Spliceosome Components Assembled on Pre-mRNA. In: Hertel, K. (eds) Spliceosomal Pre-mRNA Splicing. Methods in Molecular Biology, vol 1126. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-980-2_19
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DOI: https://doi.org/10.1007/978-1-62703-980-2_19
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-980-2
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