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

Publication ,  Journal Article
Huang, J; Chen, M; Xu, ES; Luo, L; Ma, Y; Huang, W; Floyd, W; Klann, TS; Kim, SY; Gersbach, CA; Cardona, DM; Kirsch, DG
Published in: Sci Rep
November 20, 2019

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.

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Published In

Sci Rep

DOI

EISSN

2045-2322

Publication Date

November 20, 2019

Volume

9

Issue

1

Start / End Page

17220

Location

England

Related Subject Headings

  • Signal Transduction
  • Sarcoma, Experimental
  • Proto-Oncogene Proteins p21(ras)
  • Neoplasm Proteins
  • Mutation
  • Mice, Nude
  • Mice, Inbred C57BL
  • Mice
  • Gene Expression Regulation, Neoplastic
  • Fibroblasts
 

Citation

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Huang, J., Chen, M., Xu, E. S., Luo, L., Ma, Y., Huang, W., … Kirsch, D. G. (2019). Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D. Sci Rep, 9(1), 17220. https://doi.org/10.1038/s41598-019-53572-w
Huang, Jianguo, Mark Chen, Eric S. Xu, Lixia Luo, Yan Ma, Wesley Huang, Warren Floyd, et al. “Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D.Sci Rep 9, no. 1 (November 20, 2019): 17220. https://doi.org/10.1038/s41598-019-53572-w.
Huang J, Chen M, Xu ES, Luo L, Ma Y, Huang W, et al. Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D. Sci Rep. 2019 Nov 20;9(1):17220.
Huang, Jianguo, et al. “Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D.Sci Rep, vol. 9, no. 1, Nov. 2019, p. 17220. Pubmed, doi:10.1038/s41598-019-53572-w.
Huang J, Chen M, Xu ES, Luo L, Ma Y, Huang W, Floyd W, Klann TS, Kim SY, Gersbach CA, Cardona DM, Kirsch DG. Genome-wide CRISPR Screen to Identify Genes that Suppress Transformation in the Presence of Endogenous KrasG12D. Sci Rep. 2019 Nov 20;9(1):17220.

Published In

Sci Rep

DOI

EISSN

2045-2322

Publication Date

November 20, 2019

Volume

9

Issue

1

Start / End Page

17220

Location

England

Related Subject Headings

  • Signal Transduction
  • Sarcoma, Experimental
  • Proto-Oncogene Proteins p21(ras)
  • Neoplasm Proteins
  • Mutation
  • Mice, Nude
  • Mice, Inbred C57BL
  • Mice
  • Gene Expression Regulation, Neoplastic
  • Fibroblasts