Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D.
Abstract
Cooperating gene mutations are typically required to transform normal cells enabling
growth in soft agar or in immunodeficient mice. For example, mutations in Kras and
transformation-related protein 53 (Trp53) are known to transform a variety of mesenchymal
and epithelial cells in vitro and in vivo. Identifying other genes that can cooperate
with oncogenic Kras and substitute for Trp53 mutation has the potential to lead to
new insights into mechanisms of carcinogenesis. Here, we applied a genome-wide CRISPR/Cas9
knockout screen in KrasG12D immortalized mouse embryonic fibroblasts (MEFs) to search
for genes that when mutated cooperate with oncogenic Kras to induce transformation.
We also tested if mutation of the identified candidate genes could cooperate with
KrasG12D to generate primary sarcomas in mice. In addition to identifying the well-known
tumor suppressor cyclin dependent kinase inhibitor 2A (Cdkn2a), whose alternative
reading frame product p19 activates Trp53, we also identified other putative tumor
suppressors, such as F-box/WD repeat-containing protein 7 (Fbxw7) and solute carrier
family 9 member 3 (Slc9a3). Remarkably, the TCGA database indicates that both FBXW7
and SLC9A3 are commonly co-mutated with KRAS in human cancers. However, we found that
only mutation of Trp53 or Cdkn2a, but not Fbxw7 or Slc9a3 can cooperate with KrasG12D
to generate primary sarcomas in mice. These results show that mutations in oncogenic
Kras and either Fbxw7 or Slc9a3 are sufficient for transformation in vitro, but not
for in vivo sarcomagenesis.
Type
Journal articleSubject
Science & TechnologyMultidisciplinary Sciences
Science & Technology - Other Topics
MOUSE MODEL
CANCER
P53
MUTATIONS
GROWTH
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https://hdl.handle.net/10161/19753Published Version (Please cite this version)
10.1038/s41598-019-53572-wPublication Info
Huang, Jianguo; Chen, Mark; Xu, Eric S; Luo, Lixia; Ma, Yan; Huang, Wesley; ... Kirsch,
David G (2019). Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence
of Endogenous KrasG12D. Scientific reports, 9(1). pp. 17220. 10.1038/s41598-019-53572-w. Retrieved from https://hdl.handle.net/10161/19753.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Diana Marcella Cardona
Associate Professor of Pathology
I am active in translational research involving gastrointestinal/hepatobiliary pathology
[specifically transplant related pathology (GVHD and rejection) and carcinogenesis
of the pancreas] and bone and soft tissue malignancies [imaging techniques for intraoperative
margin assessment].
Mark Chen
House Staff
Charles Gersbach
John W. Strohbehn Distinguished Professor of Biomedical Engineering
So Young Kim
Associate Research Professor in Molecular Genetics and Microbiology
I serve as Director of the Duke Functional Genomics Core Facility, where our central
mission is to provide resources for high-throughput analysis of gene function and
small molecule screens for drug discovery. Our core works with Duke investigators
to provide the expertise, infrastructure and libraries necessary for these screens
and can collaborate on all stages of the screening project, including study design,
assay optimization and data analysis. The facility also provides services for cus
David Guy Kirsch
Barbara Levine University Distinguished Professor
My clinical interests are the multi-modality care of patients with bone and soft tissue
sarcomas and developing new sarcoma therapies. My laboratory interests include utilizing
mouse models of cancer to study cancer and radiation biology in order to develop new
cancer therapies in the pre-clinical setting.
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