Characterisation of eGFP-transgenic BALB/c mouse strain established by lentiviral transgenesis
Characterisation of eGFP-transgenic BALB/c mouse strain established by lentiviral transgenesis
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DOI：
10.1007/s11248-009-9288-6
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发表时间：
2010-02-01
影响因子：
3
通讯作者：
Boesze, Zsuzsanna
中科院分区：
文献类型：
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作者：
Kvell, Krisztian;Czoempoely, Tamas;Boesze, Zsuzsanna
慢病毒技术是创造稳定转基因动物的有力工具。然而，不确定性仍然存在是否组成型启动子抵抗长期沉默。我们使用浓缩的基于HIV-1的慢病毒载体通过卵周注射来产生稳定的转基因BALB/c小鼠。在我们的载体中，eGFP表达由人EF 1 α启动子驱动。通过体内荧光成像、PCR、组织学和流式细胞术分析建立的转基因动物的eGFP表达。eGFP表达显示均匀分布，无镶嵌现象;然而，观察到eGFP表达的组织依赖性差异。直到第六代，只有一个新生儿显示eGFP失活。eGFP +转基因骨髓细胞在辐射嵌合体中有效地提供了长期的造血重建，用具有不同eGFP表达谱的eGFP +细胞再生所有骨髓来源的谱系。预期所建立的eGFP + BALB/c小鼠品系在各种免疫学实验中极其有用。
Lentiviral technology is a powerful tool for the creation of stable transgenic animals. However, uncertainties have remained whether constitutive promoters resist long-term silencing. We used concentrated HIV-1 based lentiviral vectors to create stable transgenic BALB/c mice by perivitelline injection. In our vectors eGFP expression was driven by the human EF1 alpha promoter. The established transgenic animals were analyzed for eGFP expression by in vivo fluorescence imaging, PCR, histology and flow-cytometry. eGFP expression showed even distribution without mosaicism; however, tissue-dependent differences of eGFP expression were observed. Up to the sixth generation only one newborn showed eGFP inactivation. eGFP + transgenic bone marrow cells efficiently provided long-term haemopoietic repopulation in radiation chimeras, regenerating all bone marrow-derived lineages with eGFP + cells with distinct eGFP expression profiles. The established eGFP + BALB/c mouse strain is expected to be extremely useful in various immunological experiments.