Department of Computer Science and Applied Mathematics, Weizmann Institute of Science, Rehovot, 7610001, Israel.
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, 7610001, Israel.
Nat Commun. 2019 Oct 8;10(1):4572. doi: 10.1038/s41467-019-12642-3.
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 = 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.
大多数人类基因都经过选择性剪接，从而使蛋白质组得到了极大扩展。剪接受到众多调控输入的限制，这使得我们难以从研究天然序列中推断出普遍规律。在这里，我们通过系统的序列改变创建了一个超过 32000 个剪接事件的合理设计文库，以剖析剪接调控的复杂性。测量 RNA 和蛋白质剪接异构体使我们能够研究剪接决策的因果关系，量化各种调控输入，并从序列和二级结构准确预测（R=0.73-0.85）异构体比率。通过对单个细胞进行分析，我们测量了剪接决策的细胞间变异性，并表明它可以被编码在 DNA 中，并受到调控输入的影响，这为研究剪接调控提供了一个新的单细胞视角。
