Promoter Elements of vav Drive Transgene Expression In Vivo Throughout the Hematopoietic Compartment

Abstract

To develop a method for targeting expression of genes to the full hematopoietic system, we have used transgenic mice to explore the transcriptional regulation of the vav gene, which is expressed throughout this compartment but rarely outside it. Previously, we showed that a cluster of elements surrounding its promoter could drive hematopoietic-specific expression of a bacterial lacZ reporter gene, but the expression was confined to lymphocytes and was sporadically silenced. Those limitations are ascribed here to the prokaryotic reporter gene. With a human CD4 (hCD4) cell surface reporter, the vav promoter elements drove expression efficiently and stably in virtually all nucleated cells of adult hematopoietic tissues but not notably in nonhematopoietic cell types. In multiple lines, hCD4 appeared on most, if not all, B and T lymphocytes, granulocytes, monocytes, megakaryocytes, eosinophils, and nucleated erythroid cells. Moreover, high levels appeared on both lineage-committed progenitors and the more primitive preprogenitors. In the fetus, expression was evident in erythroid cells of the definitive but not the primitive type. These results indicate that a prokaryotic sequence can inactivate a transcription unit and that the vavpromoter region constitutes a potent transgenic vector for the entire definitive hematopoietic compartment.

Tracking the Elusive Fibrocyte: Identification and Characterization of Collagen‐Producing Hematopoietic Lineage Cells During Murine Wound Healing

“…Because the vav 1 gene is ubiquitously expressed by all hematopoietic lineage cells [20,22,34,35], including during embryologic hematopoiesis [19], utilization of this gene to irreversibly label all cells within the hematopoietic lineage enabled us to definitively confirm the existence as well as characterize the natural time course of blood-derived collagen-producing cells in the wound. The GFP1/CD451/ CD11b1/Col I1 cells first identified in this work are consistent with traditional definitions of the fibrocyte [9,11], and were found to be actively recruited to the wound site.…”

Somatic mutations in murine models of leukemia and lymphoma: Disease specificity and clinical relevance

“…15,16,21,28 These models include mice that express NHD13, NP23, CA10, or Lin28b (Supporting Information Table S1) transgenes driven by Vav1 regulatory elements, which directs lineage specific expression to the hematopoietic compartment. 33 We used targeted re-sequencing to search for spontaneous somatic mutations in a set of genes known to be recurrently mutated in hematologic cancers ( Supporting Information Tables S2, S3). 34,35 This gene set included the 14 genes most commonly mutated in human AML (NPM1, FLT3, DNMT3A, IDH1, IDH2, NRAS, KRAS, RUNX1, TET2, TP53, CEBPA, WT1, PTPN11 and KIT), as well as the 4 genes most commonly mutated in T-ALL (NOTCH1, PTEN, PHF6 and FBXW7), supplemented with six additional genes less commonly mutated in human hematopoietic malignancy (ASXL1, CBL, JAK2, EZH2, MPL and GATA1).…”

MLF1IP promotes normal erythroid proliferation and is involved in the pathogenesis of polycythemia vera

“…The entire coding region of human MLF1IP was subcloned into the HS21/45-vav vector [18], which was kindly provided by Dr Jerry M. Adams (Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia). After removing the pIC19H backbone with HindIII, the linearized vector was microinjected into the pronuclei of C57BL/6J fertilized eggs to generate the founder mice.…”