Rosa26-targeted swine models for stable gene over-expression and Cre-mediated lineage tracing

Li, Xiaoping; Yang, Yi; Bu, Lei; Guo, Xiaogang; Tang, Chengcheng; Song, Jun; Fan, Nana; Zhao, Bingzi; Ouyang, Zhen; Liu, Zhaoming; Zhao, Yu; Yi, Xiaoling; Quan, Longquan; Liu, Songcai; Yang, Zhen-Guo; Ouyang, Hongsheng; Chen, Y Eugene; Zhong, Wang; Lai, Liangxue

doi:10.1038/cr.2014.15

Cited by 84 publications

(85 citation statements)

References 10 publications

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“…13,14,21 Current evidence indicates that porcine ROSA26 shares these properties. 12,22 Here, we demonstrate serial placement of single-copy transgenes, each driven by its own promoter, with high-level expression supported by this permissive locus. This is the first report of serial transgene placement at ROSA26 in any species.…”

Section: Discussionmentioning

confidence: 84%

Strong xenoprotective function by single‐copy transgenes placed sequentially at a permissive locus

Rieblinger

Fischer

Kind

et al. 2018

Xenotransplantation

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Section: Discussionmentioning

confidence: 84%

Strong xenoprotective function by single‐copy transgenes placed sequentially at a permissive locus

Rieblinger

Fischer

Kind

et al. 2018

Xenotransplantation

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“…In addition, the transgenic-cloned Huanjiang Xiang pigs generated in the present study were carrying random integration of exogenous eGFP into genome. Studies have been pointed that the transgene expression often effected with the random integrated position in genome, leading to unpredictable expression and unstable phenotypes (Li et al, 2014;Ruan et al, 2015). Now, the stable and robust overexpression of any gene of interest can be achieved by introducing the transgene into the genomic "safe harbour" loci, such as ROSA26 (Li et al, 2014) and pH11 (Ruan et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…Studies have been pointed that the transgene expression often effected with the random integrated position in genome, leading to unpredictable expression and unstable phenotypes (Li et al, 2014;Ruan et al, 2015). Now, the stable and robust overexpression of any gene of interest can be achieved by introducing the transgene into the genomic "safe harbour" loci, such as ROSA26 (Li et al, 2014) and pH11 (Ruan et al, 2015). Such strategy is worthy of reference for the authors in future to construct genetically modified Huanjiang Xiang pigs with agricultural and biomedical applications.…”

Section: Discussionmentioning

confidence: 99%

Generation of transgenic‐cloned Huanjiang Xiang pigs systemically expressing enhanced green fluorescent protein

Zhu

Zhong²,

Ge³

et al. 2018

Reprod Domestic Animals

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Huanjiang Xiang pig is a unique native minipig breed originating in Guangxi, China, and has great utility value in agriculture and biomedicine. Reproductive biotechnologies such as somatic cell nuclear transfer (SCNT) and SCNT-mediated genetic modification show great potential value in genetic preservation and utilization of Huanjiang Xiang pigs. Our previous work has successfully produced cloned and transgenic-cloned embryos using somatic cells from a Huanjiang Xiang pig. In this study, we firstly report the generation of transgenic-cloned Huanjiang Xiang pigs carrying an enhanced green fluorescent protein (eGFP) gene. A total of 504 SCNT-derived embryos were transferred to two surrogate recipients, one of which became pregnant and gave birth to three live piglets. Exogenous eGFP transgene had integrated in all of the three Huanjiang Xiang piglets identified by genotyping. Furthermore, expression of eGFP was also detected from in vitro cultured skin fibroblast cells and various organs or tissues from positive transgenic-cloned Huanjiang Xiang pigs. The present work provides a practical method to preserve this unique genetic resource and also lays a foundation for genetic modification of Huanjiang Xiang pigs with improved values in agriculture and biomedicine.

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“…This line can be used to investigate Li Fraumeni syndrome and to decipher the oncogenic characteristics of p53 mutations found in human cancers. In addition, the generation and investigation of other Cre Tg pigs has recently been reported [24,30,31]. The development of diagnoses and therapies for human diseases requires convincing animal model data.…”

Section: Discussionmentioning

confidence: 99%

Expression of Cre Recombinase in Alveolar Epithelial Cells of the AQP2-Cre Transgenic Mini-Pigs

Luo

Huang

et al. 2014

Cell Physiol Biochem

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Background/Aims: Optimal use of Cre mediated recombination in conditional animal models depends on well characterized Cre driver lines. Unfortunately, some Cre driver lines exhibit unexpected expression patterns hindering their utility in Cre/loxP systems. Thus, systematic assessment of new Cre lines is essential for generating useful Cre driver lines for future studies. Methods: Here, we describe a Cre Transgenic (Tg) mini-pig line in which the expression of Cre is directed by a 3-kb 5' fragment of the kidney-specific aquaporin 2 (AQP2); however, the AQP2-Cre Tg mini-pig line exhibits expression of Cre in alveolar epithelial cells (AECs) instead of collecting duct cells. The specificity of the AQP2-Cre plasmid was validated in vitro, and indicating that the AQP2-Cre was specifically expressed in the transfected LLC-PK1 cells. Absolute quantitative real-time PCR (qRT-PCR) and inverse PCR were performed to determine the copy numbers and integration sites of the AQP2-Cre transgene. Relative qRT-PCR was performed to evaluate variation in Cre expression levels over time. Results: Our data indicated that this AQP2-Cre Tg mini-pig line exhibits stable expression of Cre recombinase over time and in subsequent generations, even though the AQP2-Cre transgene was segregated and reduced in subsequent generations. Conclusion: Combined with our previous studies of the activity of this Cre, we conclude that this Cre Tg mini-pig line will provide a reliable tool for generating lung-specific gene targeting mini-pig models, thereby allowing the investigation of gene functions in lung development and studying the molecular mechanisms of human lung disease.

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Rosa26-targeted swine models for stable gene over-expression and Cre-mediated lineage tracing

Cited by 84 publications

References 10 publications

Strong xenoprotective function by single‐copy transgenes placed sequentially at a permissive locus

Strong xenoprotective function by single‐copy transgenes placed sequentially at a permissive locus

Generation of transgenic‐cloned Huanjiang Xiang pigs systemically expressing enhanced green fluorescent protein

Expression of Cre Recombinase in Alveolar Epithelial Cells of the AQP2-Cre Transgenic Mini-Pigs

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