Manipulation of mouse hematopoietic progenitors by specific retroviral infection.
Previous studies have identified an enhancer 3' of the scl gene that can direct transgene expression to hematopoietic progenitors and stem cells. Here we use this enhancer to restrict expression of the avian leukosis virus receptor, TVA, to hematopoietic stem cells and progenitors in bone marrow and fetal liver and demonstrate that retroviral infection can be used to specifically introduce exogenous sequences. We show that a majority of CFU-S12 multipotential progenitor cells can be transduced in vitro. Uniquely, transduction of TVA+ progenitors with a retrovirus encoding a puromycin resistance gene allows selection and expansion of a multipotential hematopoietic progenitor population that can be superinfected with high efficiency. Using this system we show for the first time that v-Myb oncoproteins expressed from avian viruses can induce a leukemic transformation in the mouse. The phenotype of the transformed cells is similar to that which is seen in the chicken and is likewise dependent on the particular structure of v-Myb. This implies that the basic mechanisms of action of mutated transcription factors in the etiology of leukemia are conserved between birds and mammals.
Murphy
George J
GJ
Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom.
Göttgens
Berthold
B
Vegiopoulos
Alexandros
A
Sanchez
Maria-José
MJ
Leavitt
Andrew D
AD
Watson
Stephen P
SP
Green
Anthony R
AR
Frampton
Jon
J
D000818:Animals; D001854:Bone Marrow Cells / Q000821:virology; D002471:Cell Transformation, Neoplastic; D002645:Chickens; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D008099:Liver / Q000196:embryology / Q000821:virology; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D011691:Puromycin / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics*; D013234:Stem Cells / Q000378:metabolism; D019076:Transgenes
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D011691:Puromycin