Conditional gene targeting in macrophages and granulocytes using LysMcre mice
Conditional gene targeting in macrophages and granulocytes using LysMcre mice
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DOI：
10.1023/a:1008942828960
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发表时间：
1999-08-01
影响因子：
3
通讯作者：
Förster, I
中科院分区：
文献类型：
--
作者：
Clausen, BE;Burkhardt, C;Förster, I
最近，通过结合基因打靶技术和定点诱变，使用噬菌体P1衍生的Cre/loxP重组系统，小鼠中的条件诱变成为可能。这种方法的通用性取决于小鼠突变体的可用性，其中重组酶Cre在适当的细胞谱系或组织中表达。在这里，我们报告的代小鼠表达Cre髓细胞由于有针对性地插入到他们的内源性M溶菌酶基因座的Cre cDNA。在携带LysMcre等位基因和两种不同loxP侧翼靶基因之一的双突变小鼠中，在成熟巨噬细胞中测定的缺失效率为83-98，在粒细胞中测定的缺失效率接近100。在与单核细胞/巨噬细胞谱系密切相关的CD 11 c（+）脾树突细胞中可检测到部分缺失（16）。相反，在尾DNA或纯化的T和B细胞中未观察到显著缺失。总之，LysMcre小鼠允许骨髓细胞中loxP侧翼靶基因的特异性和高效的Cre介导的缺失。
Conditional mutagenesis in mice has recently been made possible through the combination of gene targeting techniques and site-directed mutagenesis, using the bacteriophage P1-derived Cre/loxP recombination system. The versatility of this approach depends on the availability of mouse mutants in which the recombinase Cre is expressed in the appropriate cell lineages or tissues. Here we report the generation of mice that express Cre in myeloid cells due to targeted insertion of the cre cDNA into their endogenous M lysozyme locus. In double mutant mice harboring both the LysMcre allele and one of two different loxP-flanked target genes tested, a deletion efficiency of 83-98 was determined in mature macrophages and near 100 in granulocytes. Partial deletion (16) could be detected in CD11c(+) splenic dendritic cells which are closely related to the monocyte/macrophage lineage. In contrast, no significant deletion was observed in tail DNA or purified T and B cells. Taken together, LysMcre mice allow for both specific and highly efficient Cre-mediated deletion of loxP-flanked target genes in myeloid cells.