Site-specific recombinase-mediated genetic technology, such as inducible Cre-loxP recombination (CreER), is widely used for in vivo genetic manipulation with temporal control. The Cre-loxP technology improves our understanding on the in vivo function of specific genes in organ development, tissue regeneration, and disease progression. However, inducible CreER often remains inefficient in gene deletion. In order to improve the efficiency of gene manipulation, we generated a self-cleaved inducible CreER (sCreER) that switches inducible CreER into a constitutively active Cre by itself. We generated endocardial driver Npr3-sCreER and fibroblast driver Col1a2-sCreER, and compared them with conventional Npr3-CreER and Col1a2-CreER, respectively. For easy-to-recombine alleles such as R26-tdTomato, there was no significant difference in recombination efficiency between sCreER and the conventional CreER. However, for alleles that were relatively inert for recombination such as R26-Confetti, R26-LZLT, R26-GFP, or VEGFR2flox/flox alleles, sCreER showed a significantly higher efficiency in recombination compared with conventional CreER in endocardial cells or fibroblasts. Compared with conventional CreER, sCreER significantly enhances the efficiency of recombination to induce gene expression or gene deletion, allowing temporal yet effective in vivo genomic modification for studying gene function in specific cell lineages.

中文翻译：

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自切诱导型Cre重组酶的产生，可进行有效的时间遗传操作。

位点特异性重组酶介导的遗传技术，例如诱导性Cre-loxP重组（CreER），已广泛用于通过时间控制的体内遗传操作。Cre-loxP技术提高了我们对特定基因在器官发育，组织再生和疾病进展中的体内功能的了解。但是，可诱导的CreER在基因删除方面通常仍然效率低下。为了提高基因操作的效率，我们生成了自切割的诱导型CreER（sCreER），该酶自身将诱导型CreER转换为组成型活性Cre。我们生成了心内膜驱动程序Npr3-sCreER和成纤维细胞驱动程序Col1a2-sCreER，并将它们分别与常规Npr3-CreER和Col1a2-CreER进行了比较。对于易于重组的等位基因，例如R26-tdTomato，sCreER和常规CreER之间的重组效率没有显着差异。但是，对于重组相对惰性的等位基因，例如R26-Confetti，R26-LZLT，R26-GFP或VEGFR2flox / flox等位基因，与常规CreER相比，sCreER在心内膜细胞或成纤维细胞中的重组效率要高得多。与传统的CreER相比，sCreER显着提高了重组诱导基因表达或基因缺失的效率，从而可以进行暂时有效的体内基因组修饰，以研究特定细胞谱系中的基因功能。与常规CreER相比，sCreER在心内膜细胞或成纤维细胞中的重组效率明显更高。与传统的CreER相比，sCreER显着提高了重组诱导基因表达或基因缺失的效率，从而可以进行暂时有效的体内基因组修饰，以研究特定细胞谱系中的基因功能。与常规CreER相比，sCreER在心内膜细胞或成纤维细胞中的重组效率明显更高。与传统的CreER相比，sCreER显着提高了重组诱导基因表达或基因缺失的效率，从而可以进行暂时有效的体内基因组修饰，以研究特定细胞谱系中的基因功能。