Comparative analysis of the efficiency and specificity of myeloid-Cre deleting strains using ROSA-EYFP reporter mice.
Comparative analysis of the efficiency and specificity of myeloid-Cre deleting strains using ROSA-EYFP reporter mice.
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DOI：
10.1016/j.jim.2014.05.009
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发表时间：
2014-06
影响因子：
2.2
通讯作者：
Lowell CA
中科院分区：
文献类型：
--
作者：
Abram CL;Roberge GL;Hu Y;Lowell CA
自从1994年在小鼠中的条件基因靶向的第一个实例以来，Cre重组酶和loxP侧翼序列的使用已经成为产生组织和时间特异性基因敲除的无价技术。在文献中已经报道的表达缺失基因序列和组织特异性或时间特异性Cre重组酶的小鼠品系的数量呈指数增长。然而，这种技术的使用越来越多，突出了几个问题，可能会影响在这些小鼠模型中观察到的任何表型的解释。特别是，准确了解Cre表达的特异性在每个菌株中是至关重要的，以便得出关于特定细胞类型在观察到的表型中的作用的结论。Cre介导的缺失特异性和效率已经在文献中以许多不同的方式描述，使得这些Cre菌株之间的直接比较不可能。在这里，我们报告了将13种不同的骨髓Cre小鼠品系与ROSA-EYFP报告小鼠杂交，并平行测定了各种幼稚未刺激造血细胞中的YFP表达。通过关注髓系亚群，我们直接比较了稳态条件下这些菌株中髓系缺失的相对效率和特异性。
Since the first example of conditional gene targeting in mice in 1994, the use of Cre recombinase and loxP flanked sequences has become an invaluable technique to generate tissue and temporal specific gene knockouts. The number of mouse strains expressing floxed-gene sequences, and tissue-specific or temporal-specific Cre-recombinase that have been reported in the literature has grown exponentially. However, increased use of this technology has highlighted several problems that can impact the interpretation of any phenotype observed in these mouse models. In particular, accurate knowledge of the specificity of Cre expression in each strain is critical in order to make conclusions about the role of specific cell types in the phenotypes observed. Cre-mediated deletion specificity and efficiency has been described in many different ways in the literature, making direct comparisons between these Cre strains impossible. Here we report crossing thirteen different myeloid-Cre mouse strains to ROSA-EYFP reporter mice and assaying YFP expression in a variety of naïve unstimulated hematopoietic cells, in parallel. By focusing on myeloid subsets, we directly compare the relative efficiency and specificity of myeloid deletion in these strains under steady-state conditions.