Rosa26 Locus Supports Tissue-specific Promoter Driving Transgene Expression Specifically in Pig

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Sep 19

PMID 25232950

Citations 22

Authors

Qingran Kong

Tang Hai

Jing Ma

Tianqing Huang

Dandan Jiang

Bingteng Xie

Meiling Wu

Jiaqiang Wang

Yuran Song

Ying Wang

Yilong He

Jialu Sun

Kui Hu

Runfa Guo

Liu Wang

Qi Zhou

Yanshuang Mu

Zhonghua Liu

Affiliations

Soon will be listed here.

Abstract

Genetically modified pigs have become a popular model system in fundamental research, agricultural and biomedical applications. However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified animals with high and consistent expressing of transgene in mouse, human and rat, as it can be targeted efficiently and is not subject to gene-silencing effects. Recently, the first case of reporter gene targeting pigs in porcine Rosa26 (pRosa26) locus was reported. In the study, full sequence of pRosa26 locus was further characterized, and the pRosa26 promoter (pR26) was cloned and we evidenced that the new porcine endogenous promoter is suitable for driving transgene expression in a high and stable manner by avoiding DNA methylation. Furthermore, elongation factor 1a promoter (EF1a) -driven GFP reporter and Myostatin promoter (MyoP)-driven Follistatin (Fst) were successfully targeted into the pRosa26 locusby traditional homologous recombination (HR) strategy. EF1a showed high activity and hypomethylation at the locus. And, muscle-specific promoter MyoP was activated strictly in muscle of the pRosa26 targeted pigs, indicating Rosa26 locus supports tissue-specific promoter driving transgene expression in its own manner. The study provided further demonstration on biomedical and agricultural applications of porcine Rosa26 promoter and locus.

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