REGULATORY MODIFICATIONS OF MICRORNA AND SHORT HAIRPIN RNA PRECURSORS

Public Deposited
Analytics

Downloadable Content

Download PDF
Request Version for Screen Reader
Last Modified
  • March 19, 2019
Creator
  • Sons, Robert
    • Affiliation: School of Medicine, Department of Cell Biology and Physiology
Abstract
  • RNA-interference (RNAi) is ubiquitously used as a research tool, with dozens of candidates using this technology currently in FDA clinical trials. A particular segment of the RNAi world, short hairpin RNAs (shRNAs) seeks to enable stable, long-term reduction of a target protein. Here, we present an optimization of shRNA structure that allows increased reduction of targeted proteins as unwanted processing events are bypassed, increasing the amount of mature RNAi species created. In Chapter 1, we present current literature as it pertains to RNAi biogenesis, regulation, and optimization of shRNA effectiveness. In Chapter 2, we present our findings of extensive shRNA precursor degradation, and complete an unbiased screen for structures that bypass this degradation and optimize RNAi activity. In Chapter 3, we offer our observations of negative processing events associated with RNA Polymerase III (RNAPIII)-driven shRNAs, including but not limited to the same extensive precursor degradation. Together, these data present new details in the dysregulation of endogenous miRNA biogenesis by shRNAs, and offer guidance towards more accurate and potent engineered shRNA regulators less likely to induce off-target effects.
Date of publication
Keyword
Subject
DOI
Identifier
Resource type
Rights statement
  • In Copyright
Advisor
  • Marzluff, Bill
  • Reader, John
  • Brennwald, Patrick
  • Hammond, Scott
  • Dominski, Zbigniew
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2015
Language
Publisher
Place of publication
  • Chapel Hill, NC
Access right
  • There are no restrictions to this item.
Date uploaded
  • August 25, 2015

Relations

Parents:

This work has no parents.

Items

Thumbnail Title Date Uploaded Visibility Actions
file details Sons_unc_0153D_15646.pdf 2019-04-10 Public Download