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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.


Gene integration and expression by PhiC31 recombinase. (A) Porcine fibroblasts with the attP-blasticidin gene were generated. AttB-DNA and PhiC31 recombinase were co-transfected into the fibroblasts and recombination occurred. (B) After recombination, the fibroblast expressed eGFP. (C) Recombination was confirmed by genomic PCR. 1, control fibroblasts; 2, attP-transfected fibroblasts; 3, recombinated fibroblasts by PhiC31.
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Figure 3: Gene integration and expression by PhiC31 recombinase. (A) Porcine fibroblasts with the attP-blasticidin gene were generated. AttB-DNA and PhiC31 recombinase were co-transfected into the fibroblasts and recombination occurred. (B) After recombination, the fibroblast expressed eGFP. (C) Recombination was confirmed by genomic PCR. 1, control fibroblasts; 2, attP-transfected fibroblasts; 3, recombinated fibroblasts by PhiC31.

Mentions: PhiC31 recombinase. Unlike Cre- and Dre-recombinase with various genome engineering functions, PhiC31 recombinase can integrate the target gene into a site-specific sequence region, attP. PhiC31 protein recombines attB and attP sequences, resulting in the gene being inserted via attB-attP recombination (Fig. 3). Several mice models have been generated using the PhiC31 integrase system. In addition, a few studies using phiC31 recombinase were carried out in livestock, particularly cattle. However, few studies involving pigs have been reported to date [241]. Therefore, we investigated the possibility of attB-attP recombination by PhiC31 in porcine cells as well. As shown in Fig. 3, DNAs (a vector DNA; attB containing a fluorescence gene and a vector DNA; phiC31) were transfected in porcine fibroblasts with attP sequences, selected, and confirmed by genomic polymerase chain reaction. Producing pig models using PhiC31 recombination research is still in the preliminary stage. However, hopefully it can become a viable option for generating conditional transgenic pigs.


Transgenesis for pig models
Gene integration and expression by PhiC31 recombinase. (A) Porcine fibroblasts with the attP-blasticidin gene were generated. AttB-DNA and PhiC31 recombinase were co-transfected into the fibroblasts and recombination occurred. (B) After recombination, the fibroblast expressed eGFP. (C) Recombination was confirmed by genomic PCR. 1, control fibroblasts; 2, attP-transfected fibroblasts; 3, recombinated fibroblasts by PhiC31.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Gene integration and expression by PhiC31 recombinase. (A) Porcine fibroblasts with the attP-blasticidin gene were generated. AttB-DNA and PhiC31 recombinase were co-transfected into the fibroblasts and recombination occurred. (B) After recombination, the fibroblast expressed eGFP. (C) Recombination was confirmed by genomic PCR. 1, control fibroblasts; 2, attP-transfected fibroblasts; 3, recombinated fibroblasts by PhiC31.
Mentions: PhiC31 recombinase. Unlike Cre- and Dre-recombinase with various genome engineering functions, PhiC31 recombinase can integrate the target gene into a site-specific sequence region, attP. PhiC31 protein recombines attB and attP sequences, resulting in the gene being inserted via attB-attP recombination (Fig. 3). Several mice models have been generated using the PhiC31 integrase system. In addition, a few studies using phiC31 recombinase were carried out in livestock, particularly cattle. However, few studies involving pigs have been reported to date [241]. Therefore, we investigated the possibility of attB-attP recombination by PhiC31 in porcine cells as well. As shown in Fig. 3, DNAs (a vector DNA; attB containing a fluorescence gene and a vector DNA; phiC31) were transfected in porcine fibroblasts with attP sequences, selected, and confirmed by genomic polymerase chain reaction. Producing pig models using PhiC31 recombination research is still in the preliminary stage. However, hopefully it can become a viable option for generating conditional transgenic 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.