Promoter elements of vav drive transgene expression in vivo throughout the hematopoietic compartment.
Promoter elements of vav drive transgene expression in vivo throughout the hematopoietic compartment.
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DOI：
10.1182/blood.v94.6.1855.418k33_1855_1863
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发表时间：
1999-09
期刊：
影响因子：
20.3
通讯作者：
S. Ogilvy;Donald Metcalf;L. Gibson;M. Bath;A. W. Harris;Jerry M. Adams
中科院分区：
文献类型：
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作者：
S. Ogilvy;Donald Metcalf;L. Gibson;M. Bath;A. W. Harris;Jerry M. Adams
为了开发一种将基因表达靶向到整个造血系统的方法，我们使用转基因小鼠来探索vav基因的转录调控，该基因在整个造血系统中表达，但很少在造血系统外表达。以前，我们发现其启动子周围的一组元件可以驱动细菌lacZ报告基因的造血特异性表达，但表达仅限于淋巴细胞，并且偶尔沉默。这些限制在此归因于原核报告基因。使用人CD4（hCD4）细胞表面报告基因，vav启动子元件在成人造血组织的几乎所有有核细胞中有效且稳定地驱动表达，但在非造血细胞类型中不明显。在多个细胞系中，hCD 4出现在大多数（如果不是全部）B和T淋巴细胞、粒细胞、单核细胞、巨核细胞、嗜酸性粒细胞和有核红细胞上。此外，高水平出现在两个谱系承诺的祖细胞和更原始的preprogenitor。在胎儿中，在定形红系细胞中表达明显，但在原始型红系细胞中表达不明显。这些结果表明，原核序列可以取代一个转录单位，并且vav启动子区构成了整个永久造血区室的有效转基因载体。
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 vav promoter region constitutes a potent transgenic vector for the entire definitive hematopoietic compartment.