A recessive screen for genes regulating hematopoietic stem cells.
Identification of genes that regulate the development, self-renewal, and differentiation of stem cells is of vital importance for understanding normal organogenesis and cancer; such knowledge also underpins regenerative medicine. Here we demonstrate that chemical mutagenesis of mice combined with advances in hematopoietic stem cell reagents and genome resources can efficiently recover recessive mutations and identify genes essential for generation and proliferation of definitive hematopoietic stem cells and/or their progeny. We used high-throughput fluorescence-activated cell sorter to analyze 9 subsets of blood stem cells, progenitor cells, circulating red cells, and platelets in more than 1300 mouse embryos at embryonic day (E) 14.5. From 45 pedigrees, we recovered 6 strains with defects in definitive hematopoiesis. We demonstrate rapid identification of a novel mutation in the c-Myb transcription factor that results in thrombocythemia and myelofibrosis as proof of principal of the utility of our fluorescence-activated cell sorter-based screen. Such phenotype-driven approaches will provide new knowledge of the genes, protein interactions, and regulatory networks that underpin stem cell biology.
Papathanasiou
Peter
P
Australian Phenomics Facility, John Curtin School of Medical Research, Australian National University, Acton, ACT, Australia. peter.papathanasiou@gmail.com
Tunningley
Robert
R
Pattabiraman
Diwakar R
DR
Ye
Ping
P
Gonda
Thomas J
TJ
Whittle
Belinda
B
Hamilton
Adam E
AE
Cridland
Simon O
SO
Lourie
Rohan
R
Perkins
Andrew C
AC
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology* / Q000502:physiology*; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005808:Genes, Recessive; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D020598:Proto-Oncogene Proteins c-myb / Q000235:genetics / Q000378:metabolism
D015415:Biomarkers; D020598:Proto-Oncogene Proteins c-myb