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| Status |
Public on Sep 17, 2007 |
| Title |
Knock-in of Kras G12D in mouse MLP-29 cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
|
| Summary |
KRAS mutations are present at a high frequency in human cancers. The development of therapies targeting mutated KRAS requires cellular and animal preclinical models. We exploited adeno-associated virus-mediated homologous recombination to insert the KRAS G12D allele in the genome of mouse somatic cells. Heterozygous mutant cells displayed a constitutively active Kras protein, marked morphologic changes, increased proliferation and motility but were not transformed. On the contrary, mouse cells in which we overexpressed the corresponding KRAS cDNA were readily transformed. The levels of Kras activation in knock-in cells were comparable with those present in human cancer cells carrying the corresponding mutation. KRAS-mutated cells were compared with their wild-type counterparts by gene expression profiling, leading to the definition of a "mutated KRAS-KI signature" of 345 genes. This signature was capable of classifying mouse and human cancers according to their KRAS mutational status, with an accuracy similar or better than published Ras signatures. The isogenic cells that we have developed recapitulate the oncogenic activation of Kras occurring in cancer and represent new models for studying Kras-mediated transformation. Our results have implications for the identification of human tumors in which the oncogenic KRAS transcriptional response is activated and suggest new strategies to build mouse models of tumor progression.
Keywords: Genetic modification by homologous recombination in somatic cells
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| Overall design |
Four clones have been analyzed, two controls (WT and NEO+) and two with the Kras G12D mutation inserted by homologous recombination (KI-1 and KI-2). The expression data are averages of technical replicates.
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| |
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| Contributor(s) |
Medico E, Isella C |
| Citation(s) |
17875685 |
| |
| Submission date |
Aug 07, 2007 |
| Last update date |
May 04, 2018 |
| Contact name |
Enzo Medico |
| E-mail(s) |
enzo.medico@ircc.it
|
| Phone |
+39-011-9933234
|
| Organization name |
Candiolo Cancer Institute, University of Torino
|
| Department |
Oncology
|
| Lab |
Laboratory of Oncogenomics
|
| Street address |
Strada Prov. 142, km 3,95
|
| City |
Candiolo |
| State/province |
TO |
| ZIP/Postal code |
10060 |
| Country |
Italy |
| |
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| Platforms (2) |
| GPL4865 |
Sentrix MouseRef-8 Expression BeadChip (Target ID) |
| GPL8321 |
[Mouse430A_2] Affymetrix Mouse Genome 430A 2.0 Array |
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| Samples (12)
|
| GSM220140 |
MLP-29 Wild-type, technical replicate a |
| GSM220141 |
MLP-29 Control NEO+ , technical replicate a |
| GSM220142 |
MLP-29 KrasG12D Clone1 technical replicate a |
|
| Relations |
| BioProject |
PRJNA101949 |
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