The central goal of this project is to understand fundamental mechanisms of human pre-mRNA splicing, a required step in the expression of most eukaryotic genes, as well as the regulation of this process. Specific mechanisms through which exons and introns are correctly identified by the spliceosome will continue to be investigated. These studies will focus on intronic sequences proximal to the splice sites, and on the RNA-binding proteins that recognize them. This project will also investigate an aspect of pre-mRNA splicing fidelity, namely the mechanisms underlying suppression of cryptic splice sites that are only used in the context of mutations elsewhere in a gene. In addition, the network of protein-protein interactions of several splicing-regulatory factors will be characterized. Finaly, the mechanisms underlying the interplay between pre- mRNA splicing and nonsense-mediated mRNA decay, an RNA quality-control process, will continue to be studied. This project will rely on integrative approaches, including biochemical, molecular, proteomics, and bioinformatics techniques, as well as both cell-based and in vitro assays. In addition to obtaining new insights into basic mechanisms of gene expression, these studies will improve the understanding of numerous mutations associated with various genetic diseases, as well as facilitate correct genetic diagnosis and therapeutics development for such diseases.

Public Health Relevance

The proposed studies have broad relevance for the majority of human genetic diseases, because a high proportion of disease-causing mutations are of the type that affect mRNA splicing and stability. This project will result in a better understanding of which mutations cause defective gene expression, and how they do so. In addition, the new findings are expected to facilitate the development of targeted therapeutics to correct certain gene-expression defects.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Method to Extend Research in Time (MERIT) Award (R37)
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Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
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Cold Spring Harbor Laboratory
Cold Spring Harbor
United States
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Wong, Mandy S; Kinney, Justin B; Krainer, Adrian R (2018) Quantitative Activity Profile and Context Dependence of All Human 5' Splice Sites. Mol Cell 71:1012-1026.e3
Sinha, Rahul; Kim, Young Jin; Nomakuchi, Tomoki et al. (2018) Antisense oligonucleotides correct the familial dysautonomia splicing defect in IKBKAP transgenic mice. Nucleic Acids Res 46:4833-4844
Sheng, Lei; Wan, Bo; Feng, Pengchao et al. (2018) Downregulation of Survivin contributes to cell-cycle arrest during postnatal cardiac development in a severe spinal muscular atrophy mouse model. Hum Mol Genet 27:486-498
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Doktor, Thomas Koed; Hua, Yimin; Andersen, Henriette Skovgaard et al. (2017) RNA-sequencing of a mouse-model of spinal muscular atrophy reveals tissue-wide changes in splicing of U12-dependent introns. Nucleic Acids Res 45:395-416
Wu, Xingxing; Wang, Shu-Huei; Sun, Junjie et al. (2017) A-44G transition in SMN2 intron 6 protects patients with spinal muscular atrophy. Hum Mol Genet 26:2768-2780
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Anczuków, Olga; Krainer, Adrian R (2015) The spliceosome, a potential Achilles heel of MYC-driven tumors. Genome Med 7:107
Xiong, Hui Y; Alipanahi, Babak; Lee, Leo J et al. (2015) RNA splicing. The human splicing code reveals new insights into the genetic determinants of disease. Science 347:1254806

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