A mouse model for visualization and conditional mutations in the erythroid lineage
A mouse model for visualization and conditional mutations in the erythroid lineage
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DOI：
10.1182/blood-2003-05-1442
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发表时间：
2004-08-01
期刊：
影响因子：
20.3
通讯作者：
Klingmüller, U
中科院分区：
文献类型：
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作者：
Heinrich, AC;Pelanda, R;Klingmüller, U
血液病可由零星或遗传突变引起。然而，导致致病性的分子机制只被部分了解。Cre-loxP重组系统促进的条件性基因操作是建立小鼠模型的一种准确方法。为了能够对红系祖细胞进行鉴定和基因组操作，我们建立了一种敲入小鼠模型(ErGFPcre)，该模型表达由内源性促红细胞生成素受体(EPOR)启动子控制的改进的GFPcre融合蛋白。我们发现ErGFPcre小鼠能够鉴定出GFP阳性的红系祖细胞，并对红系血统进行高度特异的基因组操作。对GFP阳性的红系祖细胞的分析表明，从造血干细胞室到干细胞抗原-1阴性(SCA-(1-))和c-kit(高)的早期红系祖细胞的谱系发展过程中发生了转变。在造血系统中，Cre介导的重组仅限于红系祖细胞，在成人骨髓中出现的频率高达80%，在胎儿肝脏中发生的频率接近100%。在非造血组织中观察到野生型和敲入基因的差异转录活性。因此，我们的ErGFPcre小鼠模型可以促进控制非造血性EPOR表达的调控元件的识别，并有助于红系祖细胞的特性和基因组操作。
Hematologic disorders can be caused by sporadic or inherited mutations. However, the molecular mechanisms that lead to pathogenicity are only partially understood. An accurate method to generate mouse models is conditional gene manipulation facilitated by the Cre-loxP recombination system. To enable identification and genomic manipulation of erythroid progenitor cells, we established a knock-in mouse model (ErGFPcre) that expresses an improved GFPcre fusion protein controlled by the endogenous erythropoietin receptor (EpoR) promoter. We show that ErGFPcre mice enable the identification of GFP-positive erythroid progenitor cells and the highly specific genomic manipulation of the erythroid lineage. Analysis of GFP-positive erythroid progenitor cells suggests a developmental switch in lineage progression from the hematopoietic stem cell compartment to early erythroid progenitor cells that are stem cell antigen-1-negative (Sca-(1-)) and c-kit(high). Within the hematopoietic system, Cre-mediated recombination is limited to erythroid progenitor cells and occurs in the adult bone marrow at a frequency of up to 80% and in the fetal liver with an efficiency close to 100%. Differential transcriptional activity of the wild-type and the knock-in locus was observed in nonhematopoietic tissues. Thus, our ErGFPcre mouse model could promote the identification of regulatory elements controlling nonhematopoietic EpoR expression and facilitates the characterization and genomic manipulation of erythroid progenitor cells.