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Transgenesis for pig models

View Article: PubMed Central - PubMed

ABSTRACT

Animal models, particularly pigs, have come to play an important role in translational biomedical research. There have been many pig models with genetically modifications via somatic cell nuclear transfer (SCNT). However, because most transgenic pigs have been produced by random integration to date, the necessity for more exact gene-mutated models using recombinase based conditional gene expression like mice has been raised. Currently, advanced genome-editing technologies enable us to generate specific gene-deleted and -inserted pig models. In the future, the development of pig models with gene editing technologies could be a valuable resource for biomedical research.

No MeSH data available.


Dre-rox recombination in porcine cells and embryos. (A) DNA construction and PCR-detection regions. (B) With or without Dre recombinase transfection in porcine skin fibroblasts — upper without Dre, lower with Dre. (C) Validation of DNA excision by PCR. (D) Target gene expression by Dre recombinase injection into the cloned embryos from donor cells with transfection.
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Figure 2: Dre-rox recombination in porcine cells and embryos. (A) DNA construction and PCR-detection regions. (B) With or without Dre recombinase transfection in porcine skin fibroblasts — upper without Dre, lower with Dre. (C) Validation of DNA excision by PCR. (D) Target gene expression by Dre recombinase injection into the cloned embryos from donor cells with transfection.

Mentions: Dre-rox. Recently, another site-specific recombinase, Dre, was identified in P1-like phages. Like Cre, Dre recombinase recognizes the specific sequence, rox, and causes excision of the flanked gene. Although Dre recombinase has a similar structure to Cre, it does not recognize loxP sequences, indicating that there is no crossover-recombination between Cre-rox and Dre-loxP [1]. In a study, the Cre and Dre recombination were used to produce a double conditional gene expression mouse model for retinal ganglion cell labeling [30]. However, Dre-rox recombination in pigs has not yet been investigated. As a preliminary study, our research group used porcine fibroblasts and embryos to excise the flanked fluorescence gene, rox, utilizing Dre recombinase (Fig. 2). Dre-rox can be another valuable tool conditional gene regulation in pigs.


Transgenesis for pig models
Dre-rox recombination in porcine cells and embryos. (A) DNA construction and PCR-detection regions. (B) With or without Dre recombinase transfection in porcine skin fibroblasts — upper without Dre, lower with Dre. (C) Validation of DNA excision by PCR. (D) Target gene expression by Dre recombinase injection into the cloned embryos from donor cells with transfection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037292&req=5

Figure 2: Dre-rox recombination in porcine cells and embryos. (A) DNA construction and PCR-detection regions. (B) With or without Dre recombinase transfection in porcine skin fibroblasts — upper without Dre, lower with Dre. (C) Validation of DNA excision by PCR. (D) Target gene expression by Dre recombinase injection into the cloned embryos from donor cells with transfection.
Mentions: Dre-rox. Recently, another site-specific recombinase, Dre, was identified in P1-like phages. Like Cre, Dre recombinase recognizes the specific sequence, rox, and causes excision of the flanked gene. Although Dre recombinase has a similar structure to Cre, it does not recognize loxP sequences, indicating that there is no crossover-recombination between Cre-rox and Dre-loxP [1]. In a study, the Cre and Dre recombination were used to produce a double conditional gene expression mouse model for retinal ganglion cell labeling [30]. However, Dre-rox recombination in pigs has not yet been investigated. As a preliminary study, our research group used porcine fibroblasts and embryos to excise the flanked fluorescence gene, rox, utilizing Dre recombinase (Fig. 2). Dre-rox can be another valuable tool conditional gene regulation in pigs.

View Article: PubMed Central - PubMed

ABSTRACT

Animal models, particularly pigs, have come to play an important role in translational biomedical research. There have been many pig models with genetically modifications via somatic cell nuclear transfer (SCNT). However, because most transgenic pigs have been produced by random integration to date, the necessity for more exact gene-mutated models using recombinase based conditional gene expression like mice has been raised. Currently, advanced genome-editing technologies enable us to generate specific gene-deleted and -inserted pig models. In the future, the development of pig models with gene editing technologies could be a valuable resource for biomedical research.

No MeSH data available.