Tamoxifen-inducible cardiac-specific Cre transgenic mouse using VIPR2 intron
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Laboratory Animal Research
RESEARCH
Open Access
Tamoxifen-inducible cardiac-specific Cre transgenic mouse using VIPR2 intron Hyun Jung Chin, So-young Lee and Daekee Lee*
Abstract Genetically engineered mouse models through gene deletion are useful tools for analyzing gene function. To delete a gene in a certain tissue temporally, tissue-specific and tamoxifen-inducible Cre transgenic mice are generally used. Here, we generated transgenic mouse with cardiac-specific expression of Cre recombinase fused to a mutant estrogen ligand-binding domain (ERT2) on both N-terminal and C-terminal under the regulatory region of human vasoactive intestinal peptide receptor 2 (VIPR2) intron and Hsp68 promoter (VIPR2-ERT2CreERT2). In VIPR2ERT2CreERT2 transgenic mice, mRNA for Cre gene was highly expressed in the heart. To further reveal heart-specific Cre expression, VIPR2-ERT2CreERT2 mice mated with ROSA26-lacZ reporter mice were examined by X-gal staining. Results of X-gal staining revealed that Cre-dependent recombination occurred only in the heart after treatment with tamoxifen. Taken together, these results demonstrate that VIPR2-ERT2CreERT2 transgenic mouse is a useful model to unveil a specific gene function in the heart. Keywords: VIPR2 intron, ERT2CreERT2, Heart, Tamoxifen-inducible Cre transgenic mouse
Introduction The heart is the first organ to develop. It is also the most important organ for survival during embryonic development [1]. Gene knockout experiments have shown that abnormal development of the heart is the main cause of embryonic lethality [2], making it difficult to study specific gene function in the adult heart. To resolve this limitation, Cre/loxP recombination system has been developed to examine gene functions through gene deletion using adult mice [3, 4]. Such Cre/loxP-mediated conditional gene targeting can be used to study tissue-specific gene function, including genes expressed in the heart [5]. To activate Cre in a spatiotemporal manner, tamoxifen-inducible Cre/loxP recombination system requires Cre recombinase fused with a mutant form of estrogen receptor (CreERT2) which can migrate to the nucleus and induce sitespecific recombination after treatment with tamoxifen * Correspondence: [email protected] Department of Life Science, Ewha Womans University, Ewhayeodae-gil 52, Seodaemun-gu, Seoul 03760, South Korea
[6]. Leaky Cre recombinase activity irrelevant to tamoxifen treatment can be more tightly controlled by placing the mutant estrogen receptor to both ends of Cre protein such as MerCreMer [7] and ERT2CreERT2 [8]. Regulatory DNA sequences are essential to achieve specific Cre expression. However, most Cre mouse lines express Cre in more than one specific tissue, leading to misinterpretation of data due to indirect effect of Cre-mediated gene deletion in other tissues on overall phenotype. In the present study, we selected an enhancer for heart-specific gene expression using VISTA enhancer browser and experimentally tested enhancer activity in transgenic mice [9]. Among these enhancers, hs175
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