Huei San San Leong
  • The Walter and Eliza Hall Institute of Medical Research
Research fields
  • Cancer biology
Retrovirus Mediated Malignant Transformation of Mouse Embryonic Fibroblasts
Authors:  Huei San Leong and Marnie Blewitt, date: 08/05/2013, view: 10694, Q&A: 2
Cellular transformation is a widely used method to artificially induce cells to form tumours in vivo. Here, we describe the methodology for malignant transformation of mouse embryonic fibroblasts (MEFs) for transplantation into immunodeficient nude mice, as used in Leong et al. (2013). The two-step process involves: 1) down-regulation of Trp53 expression using a short hairpin RNA (shRNA); and 2) overexpression of the oncogenic HRasV12 protein. Reduction of Trp53 expression leads to cell immortalisation, and the subsequent overexpression of oncogenic HRasV12 results in malignant transformation of a cell.
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