Mouse models of CML were generated by transducing bone marrow stem and progenitor cells with retroviruses carrying BCR-ABL (chronic phase), or BCR-ABL and NUP98-HOXA9 (blast crisis phase) and transplanted into irradiated recipient mice. The development of CML was confirmed by flow cytometry and histopathology. For Msi2 knockdown experiments, lineage negative blast crisis CML cells were infected with Msi2 or control Luciferase shRNA retroviral constructs and leukemia incidence monitored. ChIP assays were performed using the myeloid leukemia cell line M1. DNA was crosslinked and immunoprecipitated with control or anti-HOXA9 antibodies and analyzed by PCR for regions of interest. CML patient samples were obtained from the Korean Leukemia Bank (Korea), the Hammersmith MRD Lab Sample Archive (United Kingdom), the Fred Hutchinson Cancer Research Center (United States) and the Singapore General Hospital (Singapore). Gene expression in human chronic and blast crisis CML was analyzed by PCR or by DNA microarrays.
Protocol detail
Mouse Models of Chronic Myeloid Leukemia
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Anti antibodies
Blast cells
Blast crisis
Bone marrow
Cancer
Cell line
Chip assays
Dna microarrays
Flow cytometry
Gene expression
Hoxa9
Human
Korean
Leukemia
Luciferase
Mice
Myeloid leukemia
Nup98
Patient
Retroviral
Shrna
Stem
Transplanted recipient
Corresponding Organization :
Other organizations : Duke University Hospital, Duke Medical Center, Hammersmith Hospital, Imperial College London, Seoul St. Mary's Hospital, Catholic University of Korea, Singapore General Hospital, Duke-NUS Medical School, Fred Hutch Cancer Center, University of Rochester
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Variable analysis
independent variables
- Transduction of bone marrow stem and progenitor cells with retroviruses carrying BCR-ABL (chronic phase), or BCR-ABL and NUP98-HOXA9 (blast crisis phase)
- Msi2 knockdown using Msi2 or control Luciferase shRNA retroviral constructs
- Development of chronic myeloid leukemia (CML)
- Leukemia incidence
- Regions of interest bound by HOXA9 (as determined by ChIP assays)
- Gene expression in human chronic and blast crisis CML
- Irradiated recipient mice
- Control Luciferase shRNA retroviral constructs
- Control antibodies used in ChIP assays
- Not specified
- Control Luciferase shRNA retroviral constructs
- Control antibodies used in ChIP assays
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