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...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2009-08, Vol.4 (8), p.e6679-e6679
Main Authors: Kong, Qingran, Wu, Meiling, Huan, Yanjun, Zhang, Li, Liu, Haiyan, Bou, Gerelchimeg, Luo, Yibo, Mu, Yanshuang, Liu, Zhonghua
Format: Article
Language:English
Subjects:
Analysis
Animal genetic engineering
Animal models
Animals
Animals, Genetically Modified
Base Sequence
Biotechnology/Bioengineering
Bisulfite
Cloning
Copy number
Cytomegalovirus
Cytomegalovirus - genetics
Deoxyribonucleic acid
Developmental Biology
Developmental Biology/Embryology
Developmental Biology/Molecular Development
DNA
DNA Methylation
Drosophila
Epigenetics
Fibroblasts
Fluorescence
Gene Dosage
Gene expression
Genes
Genetic research
Genetically modified animals
Genetics and Genomics/Animal Genetics
Genetics and Genomics/Epigenetics
Genomes
Green fluorescent protein
Green Fluorescent Proteins - genetics
Heredity
Hogs
Insects
Life sciences
Methylation
Molecular Sequence Data
Nuclear transfer
Offspring
Pigs
promoter regions
Promoter Regions, Genetic
Proteins
reporter genes
Rodents
Sequence Homology, Nucleic Acid
Sequences
Somatic cell nuclear transfer
Stability
Sulfites
Swine
Tissues
transgene copy number
Transgenes
Transgenic animals
Trends
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0006679