Pre-mRNA剪接是真核基因表达的重要调控机制，其异常会导致多种疾病，包括神经退行性疾病，癌症等。然而，关键剪接因子的缺失往往导致动物死亡，严重限制了对其体内功能的研究。我们前期鉴定了UAF-1, SFA-1, MFAP-1等剪接因子的线虫条件致死突变，最近发现抑癌相关剪接因子基因rbm-5，脊髓肌肉萎缩症基因smn-1，以及视网膜色素变性基因prp-8和prp-6都与uaf-1互作影响线虫的存活。为深入了解SMN-1, PRP-8, PRP-6和UAF-1互作的分子机制以及U2af2/UAF-1在哺乳动物中的功能，我们拟研究：1，uaf-1(n4588)改善smn-1(lf)突变表型的分子机制；2，prp-8，prp-6与uaf-1相互作用影响RNA剪接的分子机制；3，U2af2 (T145I)突变敲进小鼠的RNA剪接和胚胎发育，为RNA剪接相关神经退行性疾病的发病机制提供新的认识。

Pre-mRNA splicing is a fundamental regulatory mechanism for eukaryotic gene expression. Mutations in splicing factors can lead to severe diseases, such as neurodegenerative diseases and cancers. However, loss-of-function mutations in key splicing factors often cause lethality, which significantly impedes functional analyses of these factors. We previously isolated conditional mutations affecting the key splicing factors UAF-1/U2AF65, SFA-1/SF1 and MFAP-1. Studies along this line allowed us to find that the tumor suppressor related gene rbm-5, the spinal muscular atrophy gene smn-1, the retinitis pigmentosa-related splicing factor genes prp-8/PRP8 and prp-6/PRP6 all interact with uaf-1 to affect animal survival. To further understand how smn-1, prp-8 and prp-6 interact with uaf-1, we propose three sets of studies: 1, we will investigate the molecular mechanism underlying the suppression of smn-1(lf) defects by uaf-1(n4588); 2, we will analyze how prp-8 or prp-6 interacts with uaf-1 to affect RNA splicing and gene expression; 3, we will investigate how the U2af2 (T145I) knockin mutation (corresponding to the UAF-1(n4588) mutation in C. elegans) affects RNA splicing and embryonic development in mice. Studies proposed here will provide novel insights into the functions of these splicing factors in the pathogenesis of neurodegenerative diseases.