Novel reporter and deleter mouse strains generated using VCre/VloxP and SCre/SloxP systems, and their system specificity in mice.
Novel reporter and deleter mouse strains generated using VCre/VloxP and SCre/SloxP systems, and their system specificity in mice.
复制标题
使用 VCre/VloxP 和 SCre/SloxP 系统生成的新型报告和删除小鼠品系，及其在小鼠中的系统特异性。
DOI：
10.1007/s11248-018-0067-0
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Ito M.
中科院分区：
文献类型：
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作者：
Yoshimura Y;Ida-Tanaka M;Hiramaki T;Goto M;Kamisako T;Eto T;Yagoto M;Kawai K;Takahashi T;Nakayama M;Ito M.
Cre/loxP位点特异性重组技术是实验动物基因操作的有力工具。VCre/VloxP和SCre/SloxP是新的位点特异性重组系统，由重组酶及其特异性识别序列组成，其功能类似于Cre/loxP。以前的报道使用大肠杆菌和Oryzias latipes证明了每种重组酶与其靶位点之间存在严格的特异性; VCre/VloxP，SCre/SloxP和Cre/loxP彼此之间没有交叉反应性。在这项研究中，我们建立了四种新的敲入（KI）小鼠品系，其中VloxP-EGFP，SloxP-tdTomato，CAG-VCre和CAG-SCre基因插入ROSA 26位点。VloxP-EGFP和SloxP-tdTomato KI小鼠是在终止密码子后携带EGFP或tdTomato基因的报告小鼠，其分别被VloxP或SloxP片段floxed。CAG-VCre和CAG-SCre KI小鼠携带普遍表达的VCre或SCre基因。将这两种报告小鼠与三种不同的删除小鼠（CAG-VCre KI、CAG-SCre KI和表达Cre的转基因小鼠）杂交。通过这些配对，我们发现VCre/VloxP和SCre/SloxP系统在小鼠中的功能与Cre/loxP相似，并且重组酶对其识别序列显示出紧密的特异性。我们的研究结果表明，这些新的重组系统允许高度复杂的基因组操作，并将是有用的跟踪多个细胞谱系的命运或阐明复杂的时空调控基因表达。
DNA site-specific recombination by Cre/loxP is a powerful tool for gene manipulation in experimental animals. VCre/VloxP and SCre/SloxP are novel site-specific recombination systems, consisting of a recombinase and its specific recognition sequences, which function in a manner similar to Cre/loxP. Previous reports usingEscherichia coliandOryzias latipesdemonstrated the existence of stringent specificity between each recombinase and its target sites; VCre/VloxP, SCre/SloxP, and Cre/loxP have no cross-reactivity with each other. In this study, we established four novel knock-in (KI) mouse strains in which VloxP-EGFP, SloxP-tdTomato, CAG-VCre, and CAG-SCre genes were inserted into the ROSA26 locus. VloxP-EGFP and SloxP-tdTomato KI mice were reporter mice carrying EGFP or tdTomato genes posterior to the stop codon, which was floxed by VloxP or SloxP fragments, respectively. CAG-VCre and CAG-SCre KI mice carried VCre or SCre genes that were expressed ubiquitously. These two reporter mice were crossed with three different deleter mice, CAG-VCre KI, CAG-SCre KI, and Cre-expressing transgenic mice. Through these matings, we found that VCre/VloxP and SCre/SloxP systems were functional in mice similar to Cre/loxP, and that the recombinases showed tight specificity for their recognition sequences. Our results suggest that these novel recombination systems allow highly sophisticated genome manipulations and will be useful for tracing the fates of multiple cell lineages or elucidating complex spatiotemporal regulations of gene expression.