The cebpa +37-kb enhancer directs transgene expression to myeloid progenitors and to long-term hematopoietic stem cells

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Abstract

C/EBPα is expressed preferentially in myeloid compared with lymphoid or erythroid cells and directs myeloid lineage specification. C/EBPα is also expressed at lower levels in HSCs and in several nonhematopoietic tissues. The Cebpa gene has a conserved, 450-bp segment at +37 kb that harbors enhancer-specific epigenetic marks and is activate in a myeloid cell line. Herein, we characterize transgenic C57BL/6 mice, in which the Cebpa enhancer and 845-bp promoter regulate a hCD4 reporter. FACS analysis, in vitro colony assays, and in vivo competitive and secondary transplantation revealed that myeloid but not MEPs or lymphoid progenitors and also functional LT-HSCs are found almost exclusively in the Cebpa-hCD4+ compared with hCD4- marrow population. hCD4+ CMP yielded predominantly myeloid, whereas hCD4 CMP- generated mainly Meg/E colonies. Providing insight into control of CMP maturation, Cebpa and Pu.1 RNAs were preferentially expressed in hCD4+ CMP, Scl, Gata2, Gata1, Klf1, Ets1, and Fli1 predominated in hCD4- CMP, and Runx1, Myb, HoxA9, and Erg levels were similar in both. Cebpa-hCD4 transgene expression was lacking in multiple nonhematopoietic tissues. In summary, the +37-kb Cebpa enhancer and promoter are sufficient for marrow myeloid progenitor and LT-HSC-specific expression.

Original languageEnglish (US)
Pages (from-to)419-426
Number of pages8
JournalJournal of Leukocyte Biology
Volume96
Issue number3
DOIs
StatePublished - 2014

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Cytidine Monophosphate
Hematopoietic Stem Cells
Transgenes
Bone Marrow
Erythroid Cells
Myeloid Cells
Inbred C57BL Mouse
Epigenomics
Transgenic Mice
Transplantation
Lymphocytes
RNA
Cell Line
Population
Genes

Keywords

  • Differentiation
  • Hematopoiesis
  • Myelopoiesis

ASJC Scopus subject areas

  • Cell Biology
  • Immunology
  • Medicine(all)

Cite this

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title = "The cebpa +37-kb enhancer directs transgene expression to myeloid progenitors and to long-term hematopoietic stem cells",
abstract = "C/EBPα is expressed preferentially in myeloid compared with lymphoid or erythroid cells and directs myeloid lineage specification. C/EBPα is also expressed at lower levels in HSCs and in several nonhematopoietic tissues. The Cebpa gene has a conserved, 450-bp segment at +37 kb that harbors enhancer-specific epigenetic marks and is activate in a myeloid cell line. Herein, we characterize transgenic C57BL/6 mice, in which the Cebpa enhancer and 845-bp promoter regulate a hCD4 reporter. FACS analysis, in vitro colony assays, and in vivo competitive and secondary transplantation revealed that myeloid but not MEPs or lymphoid progenitors and also functional LT-HSCs are found almost exclusively in the Cebpa-hCD4+ compared with hCD4- marrow population. hCD4+ CMP yielded predominantly myeloid, whereas hCD4 CMP- generated mainly Meg/E colonies. Providing insight into control of CMP maturation, Cebpa and Pu.1 RNAs were preferentially expressed in hCD4+ CMP, Scl, Gata2, Gata1, Klf1, Ets1, and Fli1 predominated in hCD4- CMP, and Runx1, Myb, HoxA9, and Erg levels were similar in both. Cebpa-hCD4 transgene expression was lacking in multiple nonhematopoietic tissues. In summary, the +37-kb Cebpa enhancer and promoter are sufficient for marrow myeloid progenitor and LT-HSC-specific expression.",
keywords = "Differentiation, Hematopoiesis, Myelopoiesis",
author = "Hong Guo and Ou Ma and Friedman, {Alan David}",
year = "2014",
doi = "10.1189/jlb.2AB0314-145R",
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TY - JOUR

T1 - The cebpa +37-kb enhancer directs transgene expression to myeloid progenitors and to long-term hematopoietic stem cells

AU - Guo, Hong

AU - Ma, Ou

AU - Friedman, Alan David

PY - 2014

Y1 - 2014

N2 - C/EBPα is expressed preferentially in myeloid compared with lymphoid or erythroid cells and directs myeloid lineage specification. C/EBPα is also expressed at lower levels in HSCs and in several nonhematopoietic tissues. The Cebpa gene has a conserved, 450-bp segment at +37 kb that harbors enhancer-specific epigenetic marks and is activate in a myeloid cell line. Herein, we characterize transgenic C57BL/6 mice, in which the Cebpa enhancer and 845-bp promoter regulate a hCD4 reporter. FACS analysis, in vitro colony assays, and in vivo competitive and secondary transplantation revealed that myeloid but not MEPs or lymphoid progenitors and also functional LT-HSCs are found almost exclusively in the Cebpa-hCD4+ compared with hCD4- marrow population. hCD4+ CMP yielded predominantly myeloid, whereas hCD4 CMP- generated mainly Meg/E colonies. Providing insight into control of CMP maturation, Cebpa and Pu.1 RNAs were preferentially expressed in hCD4+ CMP, Scl, Gata2, Gata1, Klf1, Ets1, and Fli1 predominated in hCD4- CMP, and Runx1, Myb, HoxA9, and Erg levels were similar in both. Cebpa-hCD4 transgene expression was lacking in multiple nonhematopoietic tissues. In summary, the +37-kb Cebpa enhancer and promoter are sufficient for marrow myeloid progenitor and LT-HSC-specific expression.

AB - C/EBPα is expressed preferentially in myeloid compared with lymphoid or erythroid cells and directs myeloid lineage specification. C/EBPα is also expressed at lower levels in HSCs and in several nonhematopoietic tissues. The Cebpa gene has a conserved, 450-bp segment at +37 kb that harbors enhancer-specific epigenetic marks and is activate in a myeloid cell line. Herein, we characterize transgenic C57BL/6 mice, in which the Cebpa enhancer and 845-bp promoter regulate a hCD4 reporter. FACS analysis, in vitro colony assays, and in vivo competitive and secondary transplantation revealed that myeloid but not MEPs or lymphoid progenitors and also functional LT-HSCs are found almost exclusively in the Cebpa-hCD4+ compared with hCD4- marrow population. hCD4+ CMP yielded predominantly myeloid, whereas hCD4 CMP- generated mainly Meg/E colonies. Providing insight into control of CMP maturation, Cebpa and Pu.1 RNAs were preferentially expressed in hCD4+ CMP, Scl, Gata2, Gata1, Klf1, Ets1, and Fli1 predominated in hCD4- CMP, and Runx1, Myb, HoxA9, and Erg levels were similar in both. Cebpa-hCD4 transgene expression was lacking in multiple nonhematopoietic tissues. In summary, the +37-kb Cebpa enhancer and promoter are sufficient for marrow myeloid progenitor and LT-HSC-specific expression.

KW - Differentiation

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