Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D.

Huang, Jianguo

Chen, Mark

Xu, Eric S

Luo, Lixia

Ma, Yan

Huang, Wesley

Floyd, Warren

Klann, Tyler S

Kim, So Young

Gersbach, Charles A

Cardona, Diana M

Kirsch, David G

2020-01-08T20:57:15Z

2020-01-08T20:57:15Z

2019-11-20

2020-01-08T20:57:13Z

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.

10.1038/s41598-019-53572-w

2045-2322

2045-2322

https://hdl.handle.net/10161/19753

eng

Springer Science and Business Media LLC

Scientific reports

10.1038/s41598-019-53572-w

Science & Technology

Multidisciplinary Sciences

Science & Technology - Other Topics

MOUSE MODEL

CANCER

P53

MUTATIONS

GROWTH

Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D.

Journal article

Chen, Mark|0000-0001-5616-9321

Kim, So Young|0000-0002-5671-1878

Gersbach, Charles A|0000-0003-1478-4013

Cardona, Diana M|0000-0002-6877-2280

17220

1

Pratt School of Engineering

Duke

Biomedical Engineering

Duke Cancer Institute

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School of Medicine

Duke Science & Society

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Duke Innovation & Entrepreneurship

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Surgery, Surgical Sciences

Surgery

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Radiation Oncology

Molecular Genetics and Microbiology

Pathology

Published

9