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.

Agency
National Institute of Health (NIH)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM042699-24
Application #
8629753
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
Project Start
Project End
Budget Start
Budget End
Support Year
24
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Cold Spring Harbor Laboratory
Department
Type
DUNS #
City
Cold Spring Harbor
State
NY
Country
United States
Zip Code
11724
Nomakuchi, Tomoki T; Rigo, Frank; Aznarez, Isabel et al. (2016) Antisense oligonucleotide-directed inhibition of nonsense-mediated mRNA decay. Nat Biotechnol 34:164-6
Hua, Yimin; Liu, Ying Hsiu; Sahashi, Kentaro et al. (2015) Motor neuron cell-nonautonomous rescue of spinal muscular atrophy phenotypes in mild and severe transgenic mouse models. Genes Dev 29:288-97
Krainer, Adrian R (2015) Splicing: still so much to learn. RNA 21:500-1
Akerman, Martin; Fregoso, Oliver I; Das, Shipra et al. (2015) Differential connectivity of splicing activators and repressors to the human spliceosome. Genome Biol 16:119
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
Anczuków, Olga; Krainer, Adrian R (2015) The spliceosome, a potential Achilles heel of MYC-driven tumors. Genome Med 7:107
Paz, Sean; Krainer, Adrian R; Caputi, Massimo (2014) HIV-1 transcription is regulated by splicing factor SRSF1. Nucleic Acids Res 42:13812-23
Weyn-Vanhentenryck, Sebastien M; Mele, Aldo; Yan, Qinghong et al. (2014) HITS-CLIP and integrative modeling define the Rbfox splicing-regulatory network linked to brain development and autism. Cell Rep 6:1139-52
Das, Shipra; Krainer, Adrian R (2014) Emerging functions of SRSF1, splicing factor and oncoprotein, in RNA metabolism and cancer. Mol Cancer Res 12:1195-204
Rigo, Frank; Chun, Seung J; Norris, Daniel A et al. (2014) Pharmacology of a central nervous system delivered 2'-O-methoxyethyl-modified survival of motor neuron splicing oligonucleotide in mice and nonhuman primates. J Pharmacol Exp Ther 350:46-55

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