Regulatory elements of the vav gene drive transgene expression in hematopoietic stem cells from adult mice
Regulatory elements of the vav gene drive transgene expression in hematopoietic stem cells from adult mice
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DOI：
10.1016/j.exphem.2004.01.005
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发表时间：
2004-04-01
影响因子：
2.6
通讯作者：
Bueren, JA
中科院分区：
文献类型：
--
作者：
Almarza, E;Segovia, JC;Bueren, JA
Objective.先前的研究已经表明，vav原癌基因的HS 21/45启动子驱动外源转基因在小鼠造血细胞（包括克隆原性骨髓（BM）祖细胞）中的主要表达。我们研究了该启动子在成年小鼠造血干细胞区室中的活性。产生在HS 21/45启动子控制下表达非功能性人CD 4标记基因（hCD 4）的近交Ly5.1转基因小鼠。基于hCD 4表达的强度分选来自这些动物的BM细胞。然后使用来自Ly5.2小鼠的未分级BM细胞作为参考竞争者群体，评估以转基因的高、中等或低/阴性表达为特征的级分的竞争性再增殖能力（CRA）。数据显示具有低/阴性或中等hCD 4表达的BM细胞具有非常差的造血CRA。相反，具有高hCD 4表达的BM细胞的特征在于高CRA。这些观察结果在分析受体小鼠的淋巴细胞和骨髓细胞时，在初次和二次受体的短期和长期移植后得到证实。我们的研究结果表明，第一次的调节HS 21/45序列的vav基因构成了一个有效的启动子驱动转基因表达的多能造血干细胞居住在成年小鼠的BM。因此，该启动子被提议用于开发需要在造血系统细胞（包括原始造血干细胞）中限制表达外源转基因的转基因小鼠和基因治疗载体。(C)2004年国际实验血液学学会。爱思唯尔公司出版
Objective. Previous studies have shown that the HS21/45 promoter of the vav protooncogene drives a predominant expression of exogenous transgenes in mouse hematopoietic cells, including clonogenic bone marrow (BM) progenitors. We investigated the activity of this promoter in the hematopoietic stem cell compartment of adult mice.Materials and Methods. Inbred Ly5.1 transgenic mice expressing a nonfunctional human CD4 marker gene (hCD4) under the control of the HS21/45 promoter were generated. BM cells from these animals were sorted based on the intensity of hCD4 expression. Fractions characterized by high, intermediate, or low/negative expression of the transgene were then assessed for their competitive repopulation ability (CRA), using unfractionated BM cells from Ly5.2 mice as a reference competitor population.Results. Data showed that BM cells having a low/negative or intermediate expression of hCD4 had a very poor hematopoietic CRA. In contrast, BM cells with high hCD4 expression were characterized by a high CRA. These observations were confirmed in the short- and long-term posttransplantation of primary and secondary recipients when analyzing the lymphoid and myeloid cells of recipient mice.Conclusions. Our results demonstrate for the first time that the regulatory HS21/45 sequence of the vav gene constitutes an efficient promoter for driving transgene expression in multipotent hematopoietic stem cells residing in the BM of adult mice. Thus, this promoter is proposed for the development of transgenic mice and gene therapy vectors that require restricted expression of exogenous transgenes in cells of the hematopoietic system, including primitive hematopoietic stem cells. (C) 2004 International Society for Experimental Hematology. Published by Elsevier Inc.