Gupta, Riti2015-04-292015-04-292013-06https://hdl.handle.net/1794/1887139 pages. A thesis presented to the Department of Chemistry and the Clark Honors College of the University of Oregon in partial fulfillment of the requirements for degree of Bachelor of Science, Spring 2013.Mis-regulation of the alternative splicing factor muscleblind-like I (MBNL 1) plays a significant role in the disease myotonic dystrophy (OM). MBNL 1 regulates alternative splicing of genes involved in development of skeletal muscle, heart and the central nervous system. In myotonic dystrophy the lack of properly localized MBNL 1 leads to mis-splicing of many pre-mRNAs. One of the mis-spliced pre-mRNAs is the MBNLJ pre-mRNA that codes for the MBNL 1 protein (an auto-regulated event). Specifically, the mis-splicing is aberrant inclusion of exon 5 in the MBNLJ pre-mRNA. Previous work has shown that intron 4 of the MBNLJ gene is highly conserved and contains multiple MBNLI binding sites. It has been shown that a 90-nucleotide region within intron 4 is necessary for regulation by MBNL 1. This study investigates the sensitivity of the auto-regulated MBNLJ splicing event by generating MBNL 1 dose-response curves using HEK293 cells with an inducible MBNL 1 expression system. Coupling this expression system with MBNLJ constructs containing deletions of various MBNLI binding sites within the important 90 nucleotide regulation region, it was shown that a single central MBNL1 site was fundamentally more important that other sites for splicing regulation.en-USAll Rights Reserved.Muscleblind (MBNL1)RNA-binding proteinsAlternative splicingThesis / Dissertation