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Gene expression

Splicing steps aside to consider its options

Nature Reviews Genetics volume 13, page 3 (2012)Cite this article

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Tyagi and colleagues applied fluorescent hybridization probes to track splicing events; the use of up to hundreds of fluorophores amplified the signals and allowed the visualization and counting of single transcript molecules.

The authors studied a GFP transcript with two engineered introns — consisting of canonical splicing signal sequences and distinct probe-binding sites — in Chinese hamster ovary cells. They probed the 3′ untranslated region (UTR) to localize all GFP transcripts and monitored the progression of splicing through the loss of colocalization with the intronic probes. They found that one of the introns was always spliced out co-transcriptionally but that splicing of the other was uncoupled from transcription so that this intron was retained in the transcript throughout the nucleoplasm. This effect was independent of the order of the introns in the construct, indicating that uncoupling from transcription can be an intrinsic feature of an intron.

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ORIGINAL RESEARCH PAPER

  1. Vargas, D. Y. et al. Single-molecule imaging of transcriptionally coupled and uncoupled splicing. Cell 147, 1054–1065 (2011) Article

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  1. Darren J. Burgess
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Burgess, D. Splicing steps aside to consider its options. Nat Rev Genet 13, 3 (2012). https://doi.org/10.1038/nrg3137

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