ENDOGENOUS AND ENGINEERED EXPRESSION OF tRNA INTRON-DERIVED CIRCULAR RNAS

Public Deposited
Analytics

Downloadable Content

Download PDF
Request Version for Screen Reader
Last Modified
  • March 19, 2019
Creator
  • Noto, John
    • Affiliation: School of Medicine, Curriculum in Genetics and Molecular Biology
Abstract
  • Sophisticated methods of ribonucleic acid (RNA) detection including RNA-sequencing (RNA-seq) have been used to rapidly expand the known transcriptomic landscape, and formerly underappreciated RNA species including circular RNAs (circRNAs) have emerged as important subjects of research. Our lab recently developed a bioinformatics tool called Vicinal used to map RNA-seq reads that cannot be mapped end-to-end to the genome. Using a modified version of this software, we discovered abundant reads mapping to the 5’- and 3’-ends of tRNA introns in Drosophila that could best be explained by circularization of introns. We have termed these tRNA intronic circular RNAs (tricRNAs). We have used numerous assays to show that the predominant form of a particular excised intron, tric31905, is circular in animals. This tricRNA is unusual in length, displays sequence conservation among Drosophilids, and could be processed into small RNAs. We have shown that tric31905 has a dynamic expression profile during fly development, and is more abundant in certain tissues. Further research is necessary to conclude whether the cell has evolved functions for tricRNAs. Additionally, we have established that the cell uses a conserved mechanism to generate tricRNAs, and have harnessed the endogenous machinery to robustly generate stable, “designer” circRNAs in cell cultures and in living animals. We utilize a powerful imaging technique, using the aptamer sequence Broccoli as a reporter to readily track expression of engineered tricRNAs. This system could be a powerful tool for research in tRNA processing, and could be a platform for the delivery of effector RNAs, therapeutic or otherwise.
Date of publication
Keyword
DOI
Resource type
Rights statement
  • In Copyright
Advisor
  • Crews, Stephen
  • Matera, Arnold
  • Marzluff, William
  • McKay, Daniel
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2017
Language

Relations

Parents:

This work has no parents.

Items

Thumbnail Title Date Uploaded Visibility Actions
file details Noto_unc_0153D_17466.pdf 2019-04-15 Public Download