Intersectional Cre driver lines generated using split-intein mediated split-Cre reconstitution.

Intersectional Cre driver lines generated using split-intein mediated split-Cre reconstitution.
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DOI:
10.1038/srep00497
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发表时间:
2012
期刊:
影响因子:
4.6
通讯作者:
Wang, Fan
Wang, Fan
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Wang, Ping;Chen, Tianrui;Sakurai, Katsuyasu;Han, Bao-Xia;He, Zhigang;Feng, Guoping;Wang, Fan

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组织和细胞类型高度特异性的Cre驱动程序是非常罕见的,因为用于指导Cre表达的大多数基因或启动子通常在多于一种组织和/或多种细胞类型中表达。我们开发了一种基于分裂内含肽的分裂Cre系统,用于通过蛋白质剪接进行高效的Cre重建。这种分裂-内含肽-分裂-Cre系统可用于交叉两个基因或启动子的表达模式,以限制在其重叠结构域中的全长Cre重建。为了在体内测试该系统,我们选择了几种保守的人增强子来驱动Cre-N-内含肽-N或内含肽-C-Cre-C转基因在不同脑区域中的表达。在所有配对的CreN/CreC转基因小鼠中,Cre依赖的报告基因在两个增强子的交叉表达域中被有效地特异性诱导。这种基于分裂内含肽的方法与用于产生高度限制的交叉Cre驱动程序以研究复杂组织(如神经系统)的其他策略相比更容易实现。
Tissue and cell type highly specific Cre drivers are very rare due to the fact that most genes or promoters used to direct Cre expressions are generally expressed in more than one tissues and/or in multiple cell types. We developed a split-intein based split-Cre system for highly efficient Cre-reconstitution through protein splicing. This split-intein-split-Cre system can be used to intersect the expression patterns of two genes or promoters to restrict full-length Cre reconstitution in their overlapping domains. To test this system in vivo, we selected several conserved human enhancers to drive the expression of either Cre-N-intein-N, or intein-C-Cre-C transgene in different brain regions. In all paired CreN/CreC transgenic mice, Cre-dependent reporter was efficiently induced specifically in the intersectional expression domains of two enhancers. This split-intein based method is simpler to implement compared with other strategies for generating highly-restricted intersectional Cre drivers to study complex tissues such as the nervous system.
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