Back A, East K, Hickstein D
Medical Research Service, Seattle Veterans Affairs Medical Center, WA.
Blood. 1995 Feb 15;85(4):1017-24.
The human leukocyte integrin subunit CD11b is expressed predominantly on myelomonocytic cells. To identify CD11b promoter sequences important for myelomonocytic gene expression and to assess the utility of the CD11b promoter for expressing heterologous genes in vivo, we generated transgenic mice with a human CD4 reporter gene driven by CD11b promoter constructs composed of 1.5, 0.3, or 0.1 kb of DNA sequence 5' to the transcription start site. Using flow cytometry to detect the human CD4 reporter on murine leukocytes, two of three 1.5-kb CD11b promoter founder lines showed surface expression of the human CD4 transgene in granulocytes and lymphocytes. The transgene expression observed in lymphocytes was inappropriate relative to the normal pattern of CD11b expression. Of the eight 0.3-kb or 0.1-kb founder lines, only one 0.1-kb founder line showed transgene expression. The overall pattern of transgene expression among the 11 founder lines does not parallel expression of the endogenous CD11b gene. These studies indicate that additional CD11b regulatory elements will be required to express a reporter gene in vivo in a lineage-specific pattern that mimics the endogenous CD11b gene.
人类白细胞整合素亚基CD11b主要在骨髓单核细胞上表达。为了鉴定对骨髓单核细胞基因表达重要的CD11b启动子序列,并评估CD11b启动子在体内表达异源基因的效用,我们构建了转基因小鼠,其携带由转录起始位点上游1.5 kb、0.3 kb或0.1 kb DNA序列驱动的人类CD4报告基因的CD11b启动子构建体。使用流式细胞术检测小鼠白细胞上的人类CD4报告基因,三个1.5 kb CD11b启动子奠基系中的两个在粒细胞和淋巴细胞中显示出人类CD4转基因的表面表达。相对于CD11b的正常表达模式,在淋巴细胞中观察到的转基因表达是不适当的。在八个0.3 kb或0.1 kb奠基系中,只有一个0.1 kb奠基系显示出转基因表达。11个奠基系中转基因表达的总体模式与内源性CD11b基因的表达不平行。这些研究表明,需要额外的CD11b调控元件,以便以模拟内源性CD11b基因的谱系特异性模式在体内表达报告基因。