Dissecting splicing decisions and cell-to-cell variability with designed sequence libraries.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 31594945.
- Also identified by DOI 10.1038/s41467-019-12642-3 and PMC identifier 6783452.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Most human genes are alternatively spliced, allowing for a large expansion of the proteome. The multitude of regulatory inputs to splicing limits the potential to infer general principles from investigating native sequences. Here, we create a rationally designed library of >32,000 splicing events to dissect the complexity of splicing regulation through systematic sequence alterations. Measuring RNA and protein splice isoforms allows us to investigate both cause and effect of splicing decisions, quantify diverse regulatory inputs and accurately predict (R<sup>2</sup> = 0.73-0.85) isoform ratios from sequence and secondary structure. By profiling individual cells, we measure the cell-to-cell variability of splicing decisions and show that it can be encoded in the DNA and influenced by regulatory inputs, opening the door for a novel, single-cell perspective on splicing regulation.
Medical subject headings
- Alternative Splicing
- Proteome
- RNA, Messenger
- Single-Cell Analysis