The Experts below are selected from a list of 28035 Experts worldwide ranked by ideXlab platform
Hongsheng Ouyang - One of the best experts on this subject based on the ideXlab platform.
-
Germ cell-specific expression of Cre Recombinase using the VASA promoter in the pig.
FEBS open bio, 2015Co-Authors: Yuning Song, Daxin Pang, Liangxue Lai, Yongye Huang, Anfeng Wang, Xiaochun Tang, Hongsheng OuyangAbstract:The Cre-loxP system is a powerful tool for genetic analysis of distinct cell lineages and tissue-specific gene knockout in animal models. VASA is specifically expressed in reproductive tissues, and is known to play important roles in spermatogenesis and germ-cell growth. In this study, Cre Recombinase transgenic pigs under the control of the VASA promoter were generated by somatic cell nuclear transfer. Germ cell-specific expression of Cre Recombinase in VASA-Cre transgenic pigs was shown by western blotting and immunohistochemistry. VASA-Cre transgenic pigs will be a useful tool for germ cell-specific gene knockout and a disease model for disorders of the reproductive system.
-
Generation of AQP2-Cre transgenic mini-pigs specifically expressing Cre Recombinase in kidney collecting duct cells
Transgenic research, 2013Co-Authors: Weiwei Luo, Yongye Huang, Yang Han, Chaogang Yao, Xinping Duan, Hongsheng OuyangAbstract:The important differences in physiological parameters and anatomical characteristics of the kidney between humans and mice make it difficult to replicate the precise progression of human renal cystic diseases in gene modification mouse models. In contrast to mice, pigs are a better animal model of human diseases, as they are more similar in terms of organ size, structure, and physiological parameters. Here, we report the generation and initial examination of an AQP2-Cre transgenic (Tg) Chinese miniature (mini)-pig line that expresses Cre Recombinase exclusively in kidney collecting duct cells. An 8-kb fragment of the mini-pig aquaporin 2 (AQP2) 5′-flanking region was utilized to direct Cre expression in Tg mini-pigs. Two Tg mini-pigs were generated by pig somatic cell nuclear transfer and both carried the entire coding sequence of Cre Recombinase. RT-PCR and western blotting analysis revealed that Cre Recombinase was uniquely expressed in the kidney, while immunohistochemical studies located its expression in kidney collecting duct cells. Furthermore, six integration sites and 12–14 copies of the Cre gene were detected in various tissues by high-efficiency thermal asymmetric interlaced PCR and absolute quantitative real-time PCR, respectively. Combined with previous studies of Cre Recombinase activity, we believe that this AQP2-Cre Tg mini-pig line will be a useful tool to generate kidney collecting duct cell-specific gene knockout mini-pig models, thereby allowing the investigation of gene functions in kidney development and the mechanisms of human renal cystic disease.
-
Establishment of a transgenic pig fetal fibroblast reporter cell line for monitoring Cre Recombinase activity.
DNA and cell biology, 2009Co-Authors: Daxin Pang, Tiedong Wang, Limei Chen, Daibang Nie, Yan Sen, Hongsheng OuyangAbstract:The pig is considered to be the most suitable nonhuman source of organs for xenotransplantation and is widely used as a model of human disease. The Cre–LoxP system provides a powerful means of cell- or tissue-specific deletion of a targeted gene in cells or tissues of interest. Pigs expressing Cre Recombinase have a profound impact on the study of gene function and the generation of animal models of human diseases. To monitor Cre Recombinase expression in vivo, it is important to Create reporter strains. As a first step in the production of such transgenic pigs, we generated porcine fetal fibroblast cell lines conditionally expressing the gene for enhanced green fluorescent protein (EGFP). The EGFP gene is expressed only after Cre-mediated excision of LoxP-flanked stop sequences. These fetal fibroblast cell lines will be of great value for constructing reporter transgenic pigs.
Shengsong Xie - One of the best experts on this subject based on the ideXlab platform.
-
Expression of tamoxifen‐inducible Cre Recombinase in Lcn5‐CreERT2 transgenic mouse caput epididymis
Molecular reproduction and development, 2017Co-Authors: Guangxin Yao, Shengsong XieAbstract:The epididymis, which connects the testis to vas deferens, plays a crucial role regulating sperm maturation and fertilization. Here, a tamoxifen-inducible CreERT2 Recombinase transgenic mouse was generated to study the function of genes in the caput epididymis using the Cre/LoxP system, which is driven by the 1.8-kb Lcn5 promoter (Lcn5-CreERT2). Both Cre Recombinase and ERT2 mRNA were specifically expressed in the caput epididymis, beginning at postnatal Day 30 and inCreasing thereafter. Crossing these Lcn5-CreERT2 transgenic mice with Rosa26;mT/mG reporter mice, which express membrane-bound GFP (mGFP) only after Cre is active at its genetic locus, resulted in the presence of GFP only in the middle/distal caput epididymis after tamoxifen induction. Efficiency of the Cre Recombinase production in the caput epididymis was dose- and time-dependent. These tamoxifen-inducible caput epididymis-specific Cre Recombinase transgenic mice thus provides a simple approach to modulate epididymal principal cells in vivo, allowing for the genetic investigation of caput epididymis-specific gene functions during sperm maturation. This article is protected by copyright. All rights reserved
-
Lcn5 Promoter Directs the Region-Specific Expression of Cre Recombinase in Caput Epididymidis of Transgenic Mice
Biology of reproduction, 2013Co-Authors: Shengsong Xie, Guangxin Yao, Qiang Liu, Jinxiong Han, Xingxu Huang, Yonglian ZhangAbstract:Epididymis plays a crucial role in regulating the development of sperm motility and fertilizing capacity. To study the function of genes in the caput epididymidis using the Cre/loxP system, we generated Lcn5-Cre transgenic mice in which the expression of Cre Recombinase is driven by the 1.8-kb Lcn5 promoter. A total of 11 founder mice carrying the Lcn5-Cre transgene were identified by PCR from 38 offspring, and the integration efficiency was 28.9%. However, only 1 of the 11 transgenic mouse lines were revealed with the Cre Recombinase expressed specifically in caput epididymidis. Furthermore, expression of Cre mRNA was first observed on Postnatal Day 30 and continued to inCrease during development. Subsequently, Cre protein distribution was assessed by crossing Lcn5-Cre transgenic mice into the mT/mG reporter line. As expected, the Cre Recombinase activity was only found in principal cells of the middle/distal caput epididymidis. The tissue-specific expression of Cre protein in the caput epididymidis was further confirmed using Lcn5-Cre mice crossed with a mouse strain carrying Aip1 conditional alleles (Aip1(flox/+)). In summary, a transgenic mouse line expressing Cre Recombinase in principal cells of caput epididymidis was established. This transgenic mouse line can be used to generate conditional, caput epididymidis-specific knockout mouse models by crossing with mice harboring floxed (LoxP flanked) genes.
Daxin Pang - One of the best experts on this subject based on the ideXlab platform.
-
Germ cell-specific expression of Cre Recombinase using the VASA promoter in the pig.
FEBS open bio, 2015Co-Authors: Yuning Song, Daxin Pang, Liangxue Lai, Yongye Huang, Anfeng Wang, Xiaochun Tang, Hongsheng OuyangAbstract:The Cre-loxP system is a powerful tool for genetic analysis of distinct cell lineages and tissue-specific gene knockout in animal models. VASA is specifically expressed in reproductive tissues, and is known to play important roles in spermatogenesis and germ-cell growth. In this study, Cre Recombinase transgenic pigs under the control of the VASA promoter were generated by somatic cell nuclear transfer. Germ cell-specific expression of Cre Recombinase in VASA-Cre transgenic pigs was shown by western blotting and immunohistochemistry. VASA-Cre transgenic pigs will be a useful tool for germ cell-specific gene knockout and a disease model for disorders of the reproductive system.
-
Production of a reporter transgenic pig for monitoring Cre Recombinase activity
Biochemical and biophysical research communications, 2009Co-Authors: Daxin Pang, Tiedong Wang, Limei Chen, Mingjun Zhang, Na Song, Daibang Nie, Zhenwen Chen, Liangxue LaiAbstract:The pig is thought to be the most suitable non-human source of organs for xenotransplantation and is widely used as a model of human disease. Using pigs as disease models requires the design of conditional Cre Recombinase-loxP gene modifications, which, in turn, requires a Cre-expressing pig with defined patterns of expression controlled by the use of a tissue-specific promoter. In order to monitor Cre recombinant expression in vivo, it is important to Create a reporter strain. We have generated reporter a pig that is based on a single vector that drives the ubiquitous expression of the enhanced green fluorescent protein (EGFP). The EGFP gene is expressed only after Cre-mediated excision of loxP-flanked stop sequences. These reporter transgenic pigs will be of great value for monitoring Cre Recombinase activity in vivo.
-
Establishment of a transgenic pig fetal fibroblast reporter cell line for monitoring Cre Recombinase activity.
DNA and cell biology, 2009Co-Authors: Daxin Pang, Tiedong Wang, Limei Chen, Daibang Nie, Yan Sen, Hongsheng OuyangAbstract:The pig is considered to be the most suitable nonhuman source of organs for xenotransplantation and is widely used as a model of human disease. The Cre–LoxP system provides a powerful means of cell- or tissue-specific deletion of a targeted gene in cells or tissues of interest. Pigs expressing Cre Recombinase have a profound impact on the study of gene function and the generation of animal models of human diseases. To monitor Cre Recombinase expression in vivo, it is important to Create reporter strains. As a first step in the production of such transgenic pigs, we generated porcine fetal fibroblast cell lines conditionally expressing the gene for enhanced green fluorescent protein (EGFP). The EGFP gene is expressed only after Cre-mediated excision of LoxP-flanked stop sequences. These fetal fibroblast cell lines will be of great value for constructing reporter transgenic pigs.
Frank Pfrieger - One of the best experts on this subject based on the ideXlab platform.
-
Expression Patterns of Inducible Cre Recombinase Driven by Differential Astrocyte-Specific Promoters in Transgenic Mouse Lines.
Neuroscience bulletin, 2019Co-Authors: Ya-ting Chen, Qian Wang, Wei Jie, Yi-si Liu, Qiang-long You, Sophie Reibel Foisset, Frank Pfrieger, Jian-ming Yang, Tian-ming GaoAbstract:Astrocytes are the most abundant cell type in the central nervous system (CNS). They provide trophic support for neurons, modulate synaptic transmission and plasticity, and contribute to neuronal dysfunction. Many transgenic mouse lines have been generated to obtain astrocyte-specific expression of inducible Cre Recombinase for functional studies; however, the expression patterns of inducible Cre Recombinase in these lines have not been systematically characterized. We generated a new astrocyte-specific Aldh1l1-CreER
-
Expression Patterns of Inducible Cre Recombinase Driven by Differential Astrocyte-Specific Promoters in Transgenic Mouse Lines
Neuroscience bulletin, 2019Co-Authors: Ya-ting Chen, Wei Jie, Yi-si Liu, Qiang-long You, Wang Qian, Sophie Reibel, Frank PfriegerAbstract:Astrocytes are the most abundant cell type in the central nervous system (CNS). They provide trophic support for neurons, modulate synaptic transmission and plasticity, and contribute to neuronal dysfunction. Many transgenic mouse lines have been generated to obtain astrocyte-specific expression of inducible Cre Recombinase for functional studies; however, the expression patterns of inducible Cre Recombinase in these lines have not been systematically characterized. We generated a new astrocyte-specific Aldh1l1-CreERT2 knock-in mouse line and compared the expression pattern of Cre Recombinase between this and five widely-used transgenic lines (hGfap-CreERT2 from The Jackson Laboratory and The Mutant Mouse Resource and Research Center, Glast-CreERT2, Cx30-CreERT2, and Fgfr3-iCreERT2) by crossing with Ai14 mice, which express tdTomato fluorescence following Cre-mediated recombination. In adult Aldh1l1-CreERT2:Ai14 transgenic mice, tdTomato was detected throughout the CNS, and five novel morphologically-defined types of astrocyte were described. Among the six evaluated lines, the specificity of Cre-mediated recombination was highest when driven by Aldh1l1 and lowest when driven by hGfap; in the latter mice, co-staining between tdTomato and NeuN was observed in the hippocampus and cortex. Notably, evident leakage was noted in Fgfr3-iCreERT2 mice, and the expression level of tdTomato was low in the thalamus when Cre Recombinase expression was driven by Glast and in the capsular part of the central amygdaloid nucleus when driven by Cx30. Furthermore, tdTomato was clearly expressed in peripheral organs in four of the lines. Our results emphasize that the astrocyte-specific CreERT2 transgenic lines used in functional studies should be carefully selected.
Xiao Yang - One of the best experts on this subject based on the ideXlab platform.
-
Atp4b promoter directs the expression of Cre Recombinase in gastric parietal cells of transgenic mice
Journal of genetics and genomics = Yi chuan xue bao, 2010Co-Authors: Zengming Zhao, Yanxun Sun, Ning Hou, Yan Teng, Xiao YangAbstract:Parietal cells are one of the largest epithelium cells of the mucous membrane of the stomach that seCrete hydrochloric acid. To study the function of gastric parietal cells during gastric epithelium homeostasis, we generated a transgenic mouse line, namely, Atp4b-Cre, in which the expression of Cre Recombinase was controlled by a 1.0 kb promoter of mouse (-subunit of H(+)-, K(+)-ATPase gene (Atp4b). In order to test the tissue distribution and excision activity of Cre Recombinase in vivo, the Atp4b-Cre transgenic mice were bred with the reporter strain ROSA26 and a mouse strain that carries Smad4 conditional alleles (Smad4(Co/Co)). Multiple-tissue PCR of Atp4b-Cre;Smad4(Co/+) mice revealed that the recombination only happened in the stomach. As indicated by LacZ staining, ROSA26;Atp4b-Cre double transgenic mice showed efficient expression of Cre Recombinase within the gastric parietal cells. These results showed that this Atp4b-Cre mouse line could be used as a powerful tool to achieve conditional gene knockout in gastric parietal cells.
-
Capn8 promoter directs the expression of Cre Recombinase in gastric pit cells of transgenic mice
Genesis (New York N.Y. : 2000), 2009Co-Authors: Zengming Zhao, Yanxun Sun, Ning Hou, Yan Teng, Youliang Wang, Xiao YangAbstract:Gastric pit cells are high-turnover epithelial cells of the gastric mucosa. They seCrete mucus to protect the gastric epithelium from acid and pepsin. To investigate the genetic mechanisms underlying the physiological functions of gastric pit cells, we generated a transgenic mouse line, namely, Capn8-Cre, in which the expression of Cre Recombinase was controlled by the promoter of the intracellular Ca(2+)-regulated cysteine protease calpain-8. To test the tissue distribution and excision activity of Cre Recombinase, the Capn8-Cre transgenic mice were bred with the ROSA26 reporter strain and a mouse strain that carries Smad4 conditional alleles (Smad4(Co/Co)). Multiple-tissue PCR and LacZ staining demonstrated that Capn8-Cre transgenic mouse expressed Cre Recombinase in the gastric pit cells. Cre Recombinase activity was also detected in the liver and skin tissues. These data suggest that the Capn8-Cre mouse line described here could be used to dissect gene function in gastric pit cells.
-
Collagen1α1 promoter drives the expression of Cre Recombinase in osteoblasts of transgenic mice
Journal of genetics and genomics = Yi chuan xue bao, 2008Co-Authors: Lagabaiyila Zha, Ning Hou, Lin Chen, Jian Wang, Guan Yang, Yuanrong Gao, Xiao YangAbstract:Osteoblasts participate in bone formation, bone mineralization, osteoclast differentiation and many pathological processes. To study the function of genes in osteoblasts using Cre-LoxP system, we generated a mouse line expressing the Cre Recombinase under the control of the rat Collagen1alpha1 (Col1alpha1) promoter (Col1alpha1-Cre). Two founders were identified by genomic PCR from 16 offsprings, and the integration efficiency is 12.5%. In order to determine the tissue distribution and the activity of Cre Recombinase in the transgenic mice, the Col1alpha1-Cre transgenic mice were bred with the ROSA26 reporter strain and a mouse strain that carries Smad4 conditional alleles (Smad4(Co/Co)). Multiple tissue PCR of Col1alpha1-Cre;Smad4(Co/+)mice revealed the restricted Cre activity in bone tissues containing osteoblasts and tendon. LacZ staining in the Col1alpha1-Cre;ROSA26 double transgenic mice revealed that the Cre Recombinase began to express in the osteoblasts of calvaria at E14.5. Cre activity was observed in the osteoblasts and osteocytes of P10 double transgenic mice. All these data indicated that the Col1alpha1-Cre transgenic mice could serve as a valuable tool for osteoblast lineage analysis and conditional gene knockout in osteoblasts.
-
Endothelial cell-specific expression of Cre Recombinase in transgenic mice.
Yi chuan xue bao = Acta genetica Sinica, 2005Co-Authors: Xuan Cheng, Xiao-hong Tan, Jishuai Zhang, Yan-song Sun, Lin Chen, Xiao YangAbstract:Endothelial cells participate in angiogenesis, vascular homeostasis, thrombosis, inflammation and vascular wall remodeling. To study the function of genes in endothelial cells using Cre-loxP system, we generated Tie2-Cre transgenic mice, in which expression of Cre Recombinase is driven by Tie2 promoter. Total six founder mice carrying the Tie2-Cre transgene were identified by genomic PCR and Southern blot. The integration efficiency is 11%. In order to test the excision activity and tissue distribution of the Cre Recombinase, the Tie2-Cre transgenic line was crossed with the mouse strain carrying the Smad4 conditional alleles (Smad4(Co/Co)) or the reporter line ROSA26. PCR of multiple tissue DNA from Tie2-Cre; Smad4(Co/+) mice revealed the Cre activity in all tissues containing endothelial cells. We detected pan-endothelial expression of the Cre transgene in Tie2-Cre; ROSA26 double transgenic embryos by lacZ staining. Therefore, this mouse line may serve as a valuable tool for endothelial cell lineage analyses and conditional gene ablation in endothelial cells.
-
A transgenic mouse that targets the expression of Cre Recombinase in hepatocyte
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology, 2004Co-Authors: Wang Youliang, Xuan Cheng, Fang Cui, Jing Cheng, Xiao YangAbstract:OBJECTIVE: To construct a mouse that specifically expresses Cre Recombinase in hepatocyte. METHODS: A hepatocyte specific transgenic construct containing mouse albumin promoter, the Cre Recombinase gene and the poly (A) of human growth factor gene was generated. The linearized constructs were introduced into the fertilized eggs by microinjection to obtain the transgenic mice. The transcriptional specificity of Cre Recombinase was detected by reverse transcription polymerase chain reaction (RT-PCR). The expression and function of Cre Recombinase were detected by PCR and Southern Blot after crossing the Alb-Cre transgenic mice with the Smad4 conditional knockout mice. RESULTS: The linearized constructs were microinjected into 837 fertilized eggs, and then the 797 effective eggs of microinjected eggs were implanted into the oviducts of 27 pseudo pregnant mice. In the 53 offspring, there were 6 mice carrying the transgene identified by polymerase chain reaction (PCR) and Southern Blot. Cre Recombinase transcripts were detected in the livers and testis of the Alb-Cre transgenic mice using RT-PCR. The Cre Recombinase was expressed in the livers of the double heterozygous for Alb-Cre and Smad4 floxed allele, and the exon 8 floxed by loxP site was deleted. CONCLUSION: A hepatocyte-specific Cre transgenic mouse was generated successfully. The Cre Recombinase expressed specifically in liver and could mediate the recombination between loxP sites in vivo.