title: Transduction-Transplantation Mouse Model of Myeloproliferative Neoplasm.
abstract: Transduction-transplantation is a quick and efficient way to model human hematologic malignancies in mice. This technique results in expression of the gene of interest in hematopoietic cells and can be used to study the gene's role in normal and/or malignant hematopoiesis. This protocol provides a detailed description on how to perform transduction-transplantation using calreticulin (CALR) mutations recently identified in myeloproliferative neoplasm (MPN) as an example. In this protocol whole bone marrow cells from 5-flurouracil (5-FU) treated donor mice are transduced with a retrovirus encoding mutant CALR and transplanted into lethally irradiated syngeneic hosts. Donor cells expressing mutant CALR are marked with green fluorescent protein (GFP). Transplanted mice develop an MPN phenotype including elevated platelets in the peripheral blood, expansion of megakaryocytes in the bone marrow, and bone marrow fibrosis. We provide a step-by-step account of how to generate retrovirus, calculate viral titer, transduce whole bone marrow cells, and transplant into irradiated recipient mice.
authors.affiliation: Department of Medicine, Division of Hematology/Oncology, University of California, Irvine.
authors.forename: Thanh Kim
authors.initials: TK
authors.lastname: Nguyen
authors.affiliation: Department of Medicine, Division of Hematology/Oncology, University of California, Irvine.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Morse
authors.affiliation: Department of Medicine, Division of Hematology/Oncology, University of California, Irvine; agf@uci.edu.
authors.forename: Angela G
authors.initials: AG
authors.lastname: Fleischman
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000473:pathology; D001854:Bone Marrow Cells / Q000166:cytology*; D016026:Bone Marrow Transplantation; D037282:Calreticulin / Q000235:genetics*; D004195:Disease Models, Animal; D049452:Green Fluorescent Proteins / Q000235:genetics; D006410:Hematopoiesis; D006801:Humans; D064596:Isografts; D051379:Mice; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000235:genetics*; D009369:Neoplasms; D012190:Retroviridae; D014161:Transduction, Genetic
chemical_list: D037282:Calreticulin; D049452:Green Fluorescent Proteins