Efficient in vivo manipulation of mouse genomic sequences at the zygote stage
Efficient in vivo manipulation of mouse genomic sequences at the zygote stage
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DOI：
10.1073/pnas.93.12.5860
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发表时间：
1996-06-11
影响因子：
11.1
通讯作者：
Westphal, H
中科院分区：
文献类型：
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作者：
Lakso, M;Pichel, JG;Westphal, H
我们描述了在腺病毒EIIa启动子控制下携带cre转基因小鼠系，该启动子靶向cre重组酶在早期小鼠胚胎中的表达，为了评估该重组酶在发育早期切除loxp -侧翼DNA序列的能力，我们将EIIa-cre转基因小鼠与携带loxp -侧翼目标序列的两种不同小鼠系进行了繁殖：(1)携带loxP位点的单基因靶向新霉素耐药基因的菌株和(2)携带带有内部loxP位点的多个转基因拷贝的转基因品系。将这些低携带loxP的小鼠品系与EIIa-cre小鼠交配，产生的第一代后代中，loxP位点的序列已从所有被测试的组织（包括生殖细胞）中有效地删除。这些结果证明了一种在早期小鼠胚胎中可以有效地删除loxp -侧翼DNA序列的方法，并讨论了该方法的潜在应用，包括减少多拷贝转基因位点以产生单拷贝转基因系和在种系中引入各种微妙的突变。
We describe a transgenic mouse line carrying the cre transgene under the control of the adenovirus EIIa promoter that targets expression of the Cre recombinase to the early mouse embryo, To assess the ability of this recombinase to excise loxP-flanked DNA sequences at early stages of of development, we bred EIIa-cre transgenic mice to two different mouse lines carrying loxP-flanked target sequences: (i) a strain with a single gene-targeted neomycin resistance gene flanked by loxP sites and (ii) a transgenic line carrying multiple transgene copies with internal loxP sites, Mating either of these lowP-carrying mouse lines to EIIa-cre mice resulted in first generation progeny in which the loxP-flanked sequences had been efficiently deleted from all tissues tested, including the germ cells, Interbreeding of these first generation progeny resulted in efficient germ-line transmission of the deletion to subsequent generations, These results demonstrate a method by which loxP-flanked DNA sequences can,be efficiently deleted in the early mouse embryo, Potential applications of this approach are discussed, including reduction of multicopy transgene loci to produce single-copy transgenic lines and introduction of a variety of subtle mutations into the germ line.