Global alternative splicing analysis of mutations in two new spliceosome components KIN17 and PRCC in C. elegans. Global alternative splicing analysis of mutations in two new spliceosome components KIN17 and PRCC in C. elegans

Mutant alleles of KIN17 (dxbp-1) and PRCC (prcc-1) were identifed in a forward genetic screen for suppressors of cryptic splice site activation Overall design: Strains with cryptic splicing suppressor alleles in dxbp-1 ((M107I) or (K23N)) or prcc-1 ((I371F) or (null)) along with NMD mutant allele smg-4(az152) were compared to strains carrying the smg-4 mutant allele alone to identify changes in native gene alternative splicing

本研究通过针对隐蔽剪接位点激活抑制因子的正向遗传筛选，成功鉴定得到KIN17（dxbp-1）与PRCC（prcc-1）的突变等位基因。实验设计概况：将同时携带隐蔽剪接抑制突变等位基因与无义介导mRNA降解（Nonsense-mediated mRNA decay, NMD）突变等位基因smg-4(az152)的菌株分为实验组，其中实验组菌株的dxbp-1带有(M107I)或(K23N)突变、prcc-1带有(I371F)突变或为敲除型（null）；将该实验组与仅携带smg-4突变等位基因的对照组菌株进行对比，以鉴定内源基因可变剪接的变化特征。