Full metadata
Title
Keane_Spring_2023.pdf
Description
Most protein-coding mRNAs in eukaryotes must undergo a series of processing steps so they can be exported from the nucleus and translated into protein. Cleavage and polyadenylation are vital steps in this maturation process. Improper cleavage and polyadenylation results in variation in the 3′ UTR length of genes, which is a hallmark of various human diseases.
Previous data have shown that the majority of 3’UTRs of mRNAs from the nematode Caenorhabditis elegans terminate at an adenosine nucleotide, and that mutating this adenosine disrupts the cleavage reaction. It is unclear if the adenosine is included in the mature mRNA transcript or if it is cleaved off. To address this question, we are developing a novel method called the Terminal Adenosine Methylation (TAM) assay which will allow us to precisely define whether the cleavage reaction takes place upstream or downstream of this terminal adenosine. The TAM Assay utilizes the ability of the methyltransferase domain (MTD) of the human methyltransferase METTL16 to methylate the terminal adenosine of a test mRNA transcript prior to the cleavage reaction in vivo. The presence or absence of methylation at the terminal adenosine will then be identified using direct RNA sequencing. This project focuses on 1) preparing the chimeric construct that positions the MTD on the mRNA cleavage site of a test mRNA transcript, and 2) testing the functionality of this construct in vitro and developing a transgenic C. elegans strain expressing it. The TAM assay has the potential to be a valuable tool for elucidating the role of the terminal adenosine in cleavage and polyadenylation.
Date Created
2023-05
Contributors
- Keane, Sara (Author)
- Mangone, Marco (Thesis director)
- Lapinaite, Audrone (Committee member)
- Barrett, The Honors College (Contributor)
- School of Life Sciences (Contributor)
- Department of English (Contributor)
Topical Subject
Resource Type
Extent
23 pages
Copyright Statement
In Copyright
Primary Member of
Peer-reviewed
No
Open Access
No
Series
Academic Year 2022-2023
Handle
https://hdl.handle.net/2286/R.2.N.184500
System Created
- 2023-04-10 02:03:35
System Modified
- 2023-04-11 03:41:04
- 1 year 7 months ago
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