Transgene expression is associated with copy number and cytomegalovirus promoter methylation in transgenic pigs.
Transgenic animals have been used for years to study gene function, produce important proteins, and generate models for the study of human diseases. However, inheritance and expression instability of the transgene in transgenic animals is a major limitation. Copy number and promoter methylation are known to regulate gene expression, but no report has systematically examined their effect on transgene expression. In the study, we generated two transgenic pigs by somatic cell nuclear transfer (SCNT) that express green fluorescent protein (GFP) driven by cytomegalovirus (CMV). Absolute quantitative real-time PCR and bisulfite sequencing were performed to determine transgene copy number and promoter methylation level. The correlation of transgene expression with copy number and promoter methylation was analyzed in individual development, fibroblast cells, various tissues, and offspring of the transgenic pigs. Our results demonstrate that transgene expression is associated with copy number and CMV promoter methylation in transgenic pigs.
Kong
Qingran
Q
College of life science, Northeast Agricultural University of China, Harbin, People's Republic of China.
Wu
Meiling
M
Huan
Yanjun
Y
Zhang
Li
L
Liu
Haiyan
H
Bou
Gerelchimeg
G
Luo
Yibo
Y
Mu
Yanshuang
Y
Liu
Zhonghua
Z
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D003587:Cytomegalovirus / Q000235:genetics*; D019175:DNA Methylation; D018628:Gene Dosage; D049452:Green Fluorescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D019076:Transgenes
D049452:Green Fluorescent Proteins
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