Tumor Suppressor Analysis in CML

Methods Mol Biol. 2016:1465:87-94. doi: 10.1007/978-1-4939-4011-0_8.

Abstract

Retroviral models have tremendously contributed to our understanding of CML development and have been indispensable for preclinical drug testing which facilitated the implementation of a targeted therapy. The retroviral insertion of Bcr-Abl into mice that are genetically depleted for a potential tumor suppressor is a tool to test for a specific gene function in Bcr-Abl disease. Here we describe how to generate a Bcr-Abl retrovirus that is subsequently used for infection of primary murine BM cells, which are genetically depleted for a potential tumor suppressor gene. We will suggest control experiments and outline further methods that are required to allow for assessment of disease development upon tumor suppressor knockout in CML.

Keywords: Knockout cells; Retronectin; Retroviral transduction.

MeSH terms

  • Animals
  • Bone Marrow Transplantation
  • Disease Models, Animal
  • Fusion Proteins, bcr-abl / genetics*
  • Genes, Tumor Suppressor
  • Genetic Vectors / administration & dosage
  • Humans
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / pathology
  • Mice
  • Mice, Knockout
  • Retroviridae / genetics*

Substances

  • Fusion Proteins, bcr-abl