Presentation Title
Generating cell models to study the roles of ATAD2/B during breast cancer progression
Abstract
Cancer progression results from cumulative epigenetic changes that drive and maintain the malignant phenotype. In particular, the posttranslational acetylation of histone proteins plays important roles in cancer gene expression programs during all stages of tumorigenesis. ATAD2 and ATAD2B are closely related bromodomain proteins that have been shown to bind histone H4 acetyllysine marks in vitro, and they could be directly involved in chromatin remodeling. However, despite accumulating evidence that implicates ATAD2 as a ‘histone reader’ and an oncogene in several types cancers, the functional roles of ATAD2/B in tumorigenesis remains unclear. In this project, we outline the process to generate a cell system to study the functional roles of these proteins in cell-based breast cancer progression model. Accordingly, we have designed and generated a unique Homology Directed Repair template to knock-in a variety of epitope tags. Importantly, we have incorporated fluorescent selectable marker genes that can be used to select both monoallelic and biallelic stable knock-in events. As a proof-of-principle we present our strategy to modify ATAD2/B genes with C-terminal mAID degron tags that will be used to deplete expressed endogenous ATAD2/B in a cancer progression model. Overall, this approach should facilitate clonal selection from low efficiency of CRISPR/Cas9-mediated knock-ins at the endogenous gene loci. Future studies will employ these models to unravel the roles of histone readers in breast cancer progression.
Primary Faculty Mentor Name
Seth Frietze
Secondary Mentor Name
Andrew Fritz
Status
Undergraduate
Student College
College of Arts and Sciences
Program/Major
Biological Sciences, Integrated
Primary Research Category
Biological Sciences
Secondary Research Category
Health Sciences
Generating cell models to study the roles of ATAD2/B during breast cancer progression
Cancer progression results from cumulative epigenetic changes that drive and maintain the malignant phenotype. In particular, the posttranslational acetylation of histone proteins plays important roles in cancer gene expression programs during all stages of tumorigenesis. ATAD2 and ATAD2B are closely related bromodomain proteins that have been shown to bind histone H4 acetyllysine marks in vitro, and they could be directly involved in chromatin remodeling. However, despite accumulating evidence that implicates ATAD2 as a ‘histone reader’ and an oncogene in several types cancers, the functional roles of ATAD2/B in tumorigenesis remains unclear. In this project, we outline the process to generate a cell system to study the functional roles of these proteins in cell-based breast cancer progression model. Accordingly, we have designed and generated a unique Homology Directed Repair template to knock-in a variety of epitope tags. Importantly, we have incorporated fluorescent selectable marker genes that can be used to select both monoallelic and biallelic stable knock-in events. As a proof-of-principle we present our strategy to modify ATAD2/B genes with C-terminal mAID degron tags that will be used to deplete expressed endogenous ATAD2/B in a cancer progression model. Overall, this approach should facilitate clonal selection from low efficiency of CRISPR/Cas9-mediated knock-ins at the endogenous gene loci. Future studies will employ these models to unravel the roles of histone readers in breast cancer progression.