Dkk3-Cre BAC transgenic mouse line: A tool for highly efficient gene deletion in retinal progenitor cells
Dkk3-Cre BAC transgenic mouse line: A tool for highly efficient gene deletion in retinal progenitor cells
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DOI：
10.1002/dvg.20318
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发表时间：
2007-08-01
期刊：
影响因子：
1.5
通讯作者：
Furukawa, Takahisa
中科院分区：
文献类型：
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作者：
Sato, Shigeru;Inoue, Tatsuya;Furukawa, Takahisa
为了建立小鼠视网膜祖细胞中条件性基因缺失的遗传工具，我们使用细菌人工染色体（BAC）转基因产生了 Dkk3-Cre 转基因小鼠系。通过将该转基因系（称为 BAC-Dkk3-Cre）与 CAG-CAT-Z 报告系杂交来测定体内 Cre 重组效率。该 BAC-Dkk3-Cre 系在大多数视网膜祖细胞中显示出 Cre 重组酶活性。 Cre 活性从胚胎第 10.5 天 (E10.5) 起即可检测到，并且在胚胎发生期间通常仅限于视网膜。为了验证BAC-Dkk3-Cre小鼠成功规避致死率，我们生成了Otx2(flox/flox)/BAC-Dkk3-Cre(+)小鼠作为Otx2条件敲除小鼠。 Otx2(flox/flox)/BAC-Dkk3-Cre(+)小鼠存活，其视网膜显示感光细胞、双极细胞和水平细胞等成熟细胞类型标志物缺失，相反，无长突样细胞明显增多。因此，BAC-Dkk3-Cre 转基因小鼠系为生成条件敲除小鼠系提供了强大的工具，用于研究发育中视网膜中基因功能的丧失。
To establish the genetic tools for conditional gene deletion in mouse retinal progenitors, we generated a Dkk3-Cre transgenic mouse line using bacterial artificial chromosome (BAC) transgenesis. Cre recombination efficiency in vivo was assayed by crossing this transgenic line, termed BAC-Dkk3-Cre, with the CAG-CAT-Z reporter line. This BAC-Dkk3-Cre line showed Cre recombinase activity in most retinal progenitors. Cre activity was detectable from embryonic day 10.5 (E10.5) and generally restricted to the retina during embryogenesis. To verify that BAC-Dkk3-Cre mice successfully circumvented lethality, we generated Otx2(flox/flox)/BAC-Dkk3-Cre(+) mice as Otx2 conditional knockout mice. The Otx2(flox/flox)/BAC-Dkk3-Cre(+) mice were viable, and their retina showed loss of mature cell-type markers of photoreceptor cells, bipolar cells, and horizontal cells, in contrast, amacrine-like cells noticeably increased. Thus, the BAC-Dkk3-Cre transgenic mouse line provides a powerful tool for generating conditional knockout mouse lines for studying loss of gene functions in the developing retina.