Construction of transgenic swine with induced expression of Cre recombinase

Abstract

Miniature pigs have been recognized as valuable experimental animals in medical research. However, porcine models related to gene knockout of human diseases are not widely available. The objective of this study was to establish Mx1-Cre pigs using somatic cell nuclear transfer. In this study, we created transgenic pigs using somatic cell nuclear transfer (SCNT). Transfer of 210, 230, 250 and 215 zygotes to four surrogates produced 10 piglets. The Cre recombinase expression in transgenic pigs was studied using reverse transcriptase (RT)-PCR and immunohistochemistry. Mx1-Cre swine were shown to harbor the Cre gene in their genomic DNA using the PCR. In conclusion, Mx1-Cre transgenic piglets were successfully produced by SCNT. These transgenic swine, in conjunction with inducible systems for controlling Cre expression and function, are likely to have a profound impact on the study of human diseases.

Efficiency of porcine somatic cell nuclear transfer – a retrospective study of factors related to embryo recipient and embryos transferred

“…In the present study, 93% (431/462) embryo transfers belong to “additive gene transfer” and 7% (31/462) embryo transfers belong to “homologous recombination”; there is also no significant difference in PR rate between them. Additionally, the goal of our laboratory is to optimize the porcine reproductive performance and the selected transgenes were with no apparent cytotoxicity, such as Cre-inducible EGFP ( Li et al, 2009 ), Cre recombinase ( Chen et al, 2010 ) and human apolipoprotein CIII ( Wei et al, 2012 ). Therefore, it is believed that the different genetic modifications would not largely affect the statistical analysis in this study.…”

Exogenous enzymes upgrade transgenesis and genetic engineering of farm animals

“…Equivalent conditional methods for genetic engineering of livestock are not yet available. Albeit, some steps toward establishing conditional gene targeting methods in large animals, like generation of pigs with Cre-induced expression [160,161] have been undertaken, the complete conditional system has not been validated in domestic species.…”

Using the Cre/loxsystem for targeted integration into the human genome:loxFAS‐loxP pairing and delayed introduction of Cre DNA improve gene swapping efficiency

“…Cre recombinase was first recognized as a mammalian genome editing tool in 1988 [14]. Since then, adaptations have expanded the utility of the Cre/ lox system in mammalian cells beyond mouse cell lines [20, 24–26, 28, 29] to include Chinese hamster ovary (CHO) [30], flies [12], pig [31], and human cells [32]. Prior work has developed engineered variants with optimized performance in both CHO [33] and mouse cells [29].…”