An improved mouse line for cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus
An improved mouse line for cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus
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DOI：
10.1002/dvg.20218
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发表时间：
2006-07-01
期刊：
影响因子：
1.5
通讯作者：
Riethmacher, Dieter
中科院分区：
文献类型：
--
作者：
Brockschniedier, Damian;Pechmann, Yvonne;Riethmacher, Dieter
特异性消融活生物体内细胞的方法最近已经通过稳定的小鼠系得到改进和促进，所述小鼠系携带白喉毒素（DT）或白喉毒素受体的条件表达构建体，其可以分别在Cre介导的重组或白喉毒素的应用后被激活。我们最近描述了R261 acZ DT-A系（Brockschnieder et A，2004，Mol Cell Biol 24：7636-7642），其中将loxP-条件DTA等位基因引入到普遍表达的Rosa 26基因座中。该菌株允许通过将其与组织特异性Cre系杂交来消融广谱细胞类型。出乎意料的是，R26：lacZ/DT-A系的纯合子（但非杂合子）动物在多种组织中出现了一些退行性异常。这些缺陷最可能是由来自未结合的lacZf ′ OxDT-A盒的少量毒素的泄漏表达引起的。在这里，我们表明，插入一个额外的转录调控序列（牛生长激素多聚腺苷酸化信号，BPA）的lacZ开放阅读框架后，防止在纯合子小鼠的任何缺陷的形成。该修饰不影响lacZf-DTA等位基因的功能，如通过在Cre介导的重组后完全消除少突胶质细胞所例示的。新的R26：lacZbpA（flox）DTA系预期将极大地促进细胞类型消融小鼠的可靠产生。
The means to specifically ablate cells inside of a living organism have recently been improved and facilitated by stable mouse lines, carrying conditional expression constructs for diphtheria toxin (DT) or diphtheria \ toxin receptor, that could be activated upon Cremediated recombination or the application of diphtheria toxin, respectively. We have lately described the R261acZ DT-A line (Brockschnieder et A, 2004, Mol Cell Biol 24:7636-7642) in which a loxP-conditional DTA allele was introduced into the ubiquitously expressed Rosa26 locus. This strain allowed the ablation of a wide spectrum of cell types by crossing it to tissue specific Cre lines. Unexpectedly, homozygous (but not heterozygous) animals of the R26:lacZ/DT-A line developed some degenerative abnormalities in a variety of tissues. The defects were most probably caused by leaky expression of small amounts of toxin from the unrecombined lacZf'OxDT-A cassette. Here we show that insertion of an additional transcriptional regulatory sequence (bovine growth hormone polyadenylation signal, bpA) following the lacZ open reading frame prevented the formation of any defects in homozygous mice. The modification did not affect the functionality of the lacZf"DTA allele, as exemplified by the complete ablation of oligodendrocytes upon Cremediated recombination. The novel R26:lacZbpA(flox)DTA line is expected to greatly facilitate the reliable generation of cell type ablated mice.