Co-transcriptional splicing at nucleotide resolution
Co-transcriptional splicing at nucleotide resolution"
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The sequencing methods provide single-molecule information at nucleotide resolution on the progression of splicing as a function of Pol II position along the gene. In single-molecule intron
tracking (SMIT), nascent RNA is purified and a linker is ligated to the 3′ ends of the RNA molecules to mark the position of Pol II, followed by PCR of the nascent RNA using exon-specific
5′-end primers and massively parallel paired-end sequencing. The authors performed SMIT on 87 intron-containing genes in budding yeast and found that a substantial fraction of their
transcripts were already spliced when Pol II was positioned just downstream of 3′ splice sites (3′SS). This was unexpected, as previously published data indicated that splicing might occur
when Pol II is ∼1,000 nt downstream of introns.
onset of splicing ... was calculated to occur when Pol II was only 26 ±1 nt downstream of the 3′SS
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