Developing a Simple Titration Cell-Based Splicing Assay
| dc.contributor.author | Gacke, Jake | |
| dc.date.accessioned | 2015-08-10T18:47:50Z | |
| dc.date.available | 2015-08-10T18:47:50Z | |
| dc.date.issued | 2015-06 | |
| dc.description | 38 pages. A thesis presented to the Department of Chemistry and Biochemistry and the Clark Honors College of the University of Oregon in partial fulfillment of the requirements for degree of Bachelor of Science, Spring 2015. | en_US |
| dc.description.abstract | Muscleblind-like 1 (MBNL1) is a protein that regulates the expression of genes at the step known as pre-mRNA splicing or alternative splicing. Cells use alternative splicing to generate multiple mRNAs (and proteins) from a single gene. MBNL1-mediated splicing is dependent on MBNL1’s ability to bind pre-mRNA targets, which is possible through two critical pairs of tandem zinc finger RNA binding motifs. However, these two distinct RNA binding domains (zinc fingers 1-2 and 3-4) have non-equivalent splicing function in MBNL1 dependent alternative splicing events. The zinc finger pair (1-2) has been shown to contribute significantly higher RNA binding affinity and splicing activity for several RNA substrates compared to the zinc finger pair (3-4). Therefore, a splicing factor with greater splicing activity might be constructed by increasing MBNL1’s affinity through the creation of synthetic MBNL1 containing two copies of zinc finger pair (1-2). To test this hypothesis, an MBNL1 variant replacing the zinc finger pair (3-4) domain with another copy of zinc finger pair (1-2) domain was created – MBNL1 (1-2, 1-2). A simple cell-based splicing assay was developed to compare the splicing activities of WT-MBNL1 and MBNL1 (1-2, 1-2). Preliminary results showed that MBNL1 (1-2, 1-2) had similar splicing activity compared to the wild type MBNL1. | en_US |
| dc.identifier.uri | https://hdl.handle.net/1794/19061 | |
| dc.language.iso | en_US | en_US |
| dc.publisher | University of Oregon | en_US |
| dc.rights | Creative Commons BY-NC-ND 4.0-US | en_US |
| dc.subject | Biochemistry | en_US |
| dc.subject | Alternative splicing | en_US |
| dc.subject | MBNLI | en_US |
| dc.subject | Splicing assay | en_US |
| dc.subject | Muscleblind-like 1 | en_US |
| dc.title | Developing a Simple Titration Cell-Based Splicing Assay | en_US |
| dc.type | Thesis / Dissertation | en_US |