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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 expression by cassette exchange via cyclic recombinase (Cre). (A) Floxed blasticidin-resistant gene by loxP and lox2272 were integrated into porcine cells. (B) Donor DNA (puromycin-linked RFP gene) and Cre recombinase were co-transfected and blasticidin gene was then exchanged. (C) Genomic polymerase chain reaction (PCR) on recombinant target genes confirmed cassette exchange by Cre recombinase. 1, DNA ladder; 2, wild type cells; 3, blasticidin integrated cells; 4, cassette exchanged cells; (−), negative control.
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Figure 1: Gene expression by cassette exchange via cyclic recombinase (Cre). (A) Floxed blasticidin-resistant gene by loxP and lox2272 were integrated into porcine cells. (B) Donor DNA (puromycin-linked RFP gene) and Cre recombinase were co-transfected and blasticidin gene was then exchanged. (C) Genomic polymerase chain reaction (PCR) on recombinant target genes confirmed cassette exchange by Cre recombinase. 1, DNA ladder; 2, wild type cells; 3, blasticidin integrated cells; 4, cassette exchanged cells; (−), negative control.

Mentions: Recently, transgenic pig research has been carried out using the Cre-loxP system [8202122]. In those studies, gene excision and insertion were successfully completed using Cre recombinase. Additionally, our study confirmed the viability of using Cre recombinase to execute gene cassette exchanges (Fig. 1). If transgenic pigs can be generated via SCNT using cassette exchangeable donor cells, then various genetic functions with no change in expression level can be analyzed after gene exchange.


Transgenesis for pig models
Gene expression by cassette exchange via cyclic recombinase (Cre). (A) Floxed blasticidin-resistant gene by loxP and lox2272 were integrated into porcine cells. (B) Donor DNA (puromycin-linked RFP gene) and Cre recombinase were co-transfected and blasticidin gene was then exchanged. (C) Genomic polymerase chain reaction (PCR) on recombinant target genes confirmed cassette exchange by Cre recombinase. 1, DNA ladder; 2, wild type cells; 3, blasticidin integrated cells; 4, cassette exchanged cells; (−), negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Gene expression by cassette exchange via cyclic recombinase (Cre). (A) Floxed blasticidin-resistant gene by loxP and lox2272 were integrated into porcine cells. (B) Donor DNA (puromycin-linked RFP gene) and Cre recombinase were co-transfected and blasticidin gene was then exchanged. (C) Genomic polymerase chain reaction (PCR) on recombinant target genes confirmed cassette exchange by Cre recombinase. 1, DNA ladder; 2, wild type cells; 3, blasticidin integrated cells; 4, cassette exchanged cells; (−), negative control.
Mentions: Recently, transgenic pig research has been carried out using the Cre-loxP system [8202122]. In those studies, gene excision and insertion were successfully completed using Cre recombinase. Additionally, our study confirmed the viability of using Cre recombinase to execute gene cassette exchanges (Fig. 1). If transgenic pigs can be generated via SCNT using cassette exchangeable donor cells, then various genetic functions with no change in expression level can be analyzed after gene exchange.

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.