DNA methyltransferase 1 is essential for and uniquely regulates hematopoietic stem and progenitor cells.
Dnmt1 conditional KO HSCs and progenitors
Transcription Profiling (Microarray)
Cell Type/Cell Line, Genetic Characteristics
To examine the role of Dnmt1 in adult hematopoietic stem cells (HSCs), we conditionally disrupted Dnmt1 in the hematopoietic system. Dnmt1 was conditionally deleted by injections of poly(I)-poly(C) to induce Cre expression from the Mx-Cre transgene. Control mice were also injected with poly(I)-poly(C) but do not carry the Mx-Cre transgene. Four days after completion of poly(I)-poly(C) injections, bone marrow was harvested from the mice, antibody-mediated magnetic bead selection was used to remove cells expressing mature lineage markers, and the resulting lineage-depleted cells were stained with fluorochrome-conjugated antibodies against the surface receptors c-Kit, Sca-1 and CD34. Populations of LT-HSCs, MPPs and myeloid progenitors were FACS sorted, RNA was extracted and amplified from these sorted populations and hybridized to Affymetrix microarray chips to compare changes in gene expression induced by conditional knockout of Dnmt1 compared to control in each of the three cell populations. There are 2 biological replicates for the LT-HSCs and MPPs, and 3 biological replicates for the myeloid progenitors.
Study metadata (ISA-Tab: isa_7314_593643.zip)