The pu. 1 promoter drives myeloid gene expression in zebrafish
The pu. 1 promoter drives myeloid gene expression in zebrafish
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DOI：
10.1182/blood-2003-09-3105
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发表时间：
2004-09-01
期刊：
影响因子：
20.3
通讯作者：
Look, AT
中科院分区：
文献类型：
--
作者：
Hsu, K;Traver, D;Look, AT
PU.1 是转录因子 Ets 家族的成员，在骨髓细胞和淋巴细胞的发育中发挥重要作用。为了检查斑马鱼发育过程中血细胞亚群中斑马鱼 pu.1 (zpu.1) 的表达，我们将 zpu.1 起始密码子上游的 9 kb 斑马鱼基因组片段与绿色荧光蛋白 (GFP) 连接起来，并显微注射该构建体以生成稳定的转基因系。受精后 (hpf) 16 至 28 小时，观察到 GFP 阳性荧光骨髓前体从活胚胎的前外侧中胚层迁移，其模式与内源性 zpu.1 mRNA 的表达模式重叠。对幼虫组织切片的分析揭示了发育中的斑马鱼肾脏中表达 GFP 的造血细胞。对成人全肾骨髓细胞进行流式细胞术分析，发现 2 个离散的 GFP 阳性细胞亚群，在细胞分选后表现出骨髓或早期淋巴形态。因此，此处使用的斑马鱼 zpu.1 启动子片段能够驱动胚胎和成年造血细胞亚群中的报告基因表达。这些转基因系将有助于剖析骨髓细胞分化的细胞和分子控制，并且该启动子片段可能在急性髓系白血病斑马鱼模型的开发中有用。 (C) 2004 年，美国血液学会。
PU.1 is a member of the Ets family of transcription factors and plays an essential role in the development of both myeloid and lymphoid cells. To examine zebrafish pu.1 (zpu.1) expression in sub-populations of blood cells during zebrafish development, we linked a 9-kb zebrafish genomic fragment upstream of the zpu.1 initiator codon to green fluorescent protein (GFP) and microinjected this construct to generate stable transgenic lines. GFP-positive fluorescent myeloid precursors were observed migrating from the anterolateral mesoderm in living embryos from 16 to 28 hours after fertilization (hpf) in a pattern that overlaps the expression pattern of endogenous zpu.1 mRNA. Analysis of larval histologic sections revealed GFP-expressing hematopoietic cells in the developing zebrafish kidney. Flow cytometric analysis of cells from adult whole kidney marrow revealed 2 discrete subpopulations of GFP-positive cells, which after cell sorting exhibited either myeloid or early lymphold morphology. Thus, the zebrafish zpu.1 promoter fragment used here is capable of driving reporter gene expression in subsets of embryonic and adult hematopoietic cells. These transgenic lines will be useful to dissect the cellular and molecular control of myeloid cell differentiation, and this promoter fragment may prove useful in the development of zebrafish models of acute myeloid leukemia. (C) 2004 by The American Society of Hematology.