DOI: https://doi.org/10.15368/theses.2013.88
Available at: https://digitalcommons.calpoly.edu/theses/1052
Date of Award
6-2013
Degree Name
MS in Agriculture - Animal Science
Department/Program
Animal Science
Advisor
Matt Burd
Abstract
The deregulation of the cell cycle is important in the leukemic transformation and in the continued discovery of more efficient targeted therapies. After elucidation of the role of the RNA editor ADAR1 in the malignant reprogramming of Chronic Myeloid Leukemia (CML) progenitors to leukemia stem cells (LSCs) during the chronic phase to blast crisis stages of the disease, current research seeks to uncover the mechanism of action of ADAR1 in this process. To study the potential role ADAR1 plays in the changes to cell cycle during this disease progression, a novel Fucci (fluorescent ubiquitination-based cell cycle indicator) system was introduced into the CML cell line K562 through stable transduction of both Fucci vectors. This allows for the visualization of the cell cycle via FACS (fluorescence activated cell sorting), and fluorescent live imaging. This system was validated for use in therapeutic CML research through preliminary evaluation of ADAR1 activity in specific cell cycle stages as well as the effects of a potentially novel CML therapy, 8-azaadenosine, on the cell cycle. Completion of this work shows successful creation of a novel generation of Fucci cell line; which will aid in the clarification of the changes which occur in the cell cycle during CML disease progression, as well as the effects of CML targeted therapies on cell cycle regulation though quantitative and qualitative analysis.