Chromosomal transposition of PiggyBac in mouse embryonic stem cells

Transposon systems are widely used for generating mutations in various model organisms. PiggyBac (PB) has recently been shown to transpose efficiently in the mouse germ line and other mammalian cell lines. To facilitate PB's application in mammalian genetics, we characterized the properties of...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 105; no. 27; pp. 9290 - 9295
Main Authors Wang, Wei, Lin, Chengyi, Lu, Dong, Ning, Zeming, Cox, Tony, Melvin, David, Wang, Xiaozhong, Bradley, Allan, Liu, Pentao
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 08.07.2008
National Acad Sciences
Subjects
Animals
Base Sequence
Biological Sciences
Cell lines
Chromosomes
Chromosomes, Mammalian - metabolism
Comparative analysis
DNA
DNA Methylation
DNA Transposable Elements - genetics
Embryonic Stem Cells - metabolism
Genetic loci
Genetic transposition
Genetics
Genome - genetics
Genomes
Genomics
Hybridity
Mice
Mutagenesis
Mutagens
Mutation
Plasmids
Proteins - genetics
RNA, Untranslated
Rodents
Stem cells
Transposases - metabolism
Transposons
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Summary:Transposon systems are widely used for generating mutations in various model organisms. PiggyBac (PB) has recently been shown to transpose efficiently in the mouse germ line and other mammalian cell lines. To facilitate PB's application in mammalian genetics, we characterized the properties of the PB transposon in mouse embryonic stem (ES) cells. We first measured the transposition efficiencies of PB transposon in mouse embryonic stem cells. We next constructed a PB/SB hybrid transposon to compare PB and Sleeping Beauty (SB) transposon systems and demonstrated that PB transposition was inhibited by DNA methylation. The excision and reintegration rates of a single PB from two independent genomic loci were measured and its ability to mutate genes with gene trap cassettes was tested. We examined PB's integration site distribution in the mouse genome and found that PB transposition exhibited local hopping. The comprehensive information from this study should facilitate further exploration of the potential of PB and SB DNA transposons in mammalian genetics.
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Author contributions: W.W. and P.L. designed research; W.W., C.L., D.L., Z.N., T.C., D.M., X.W., and P.L. performed research; W.W., C.L., and X.W. contributed new reagents/analytic tools; W.W., D.L., Z.N., T.C., D.M., A.B., and P.L. analyzed data; and W.W., A.B., and P.L. wrote the paper.
Edited by Kathryn V. Anderson, Sloan–Kettering Institute, New York, NY, and approved April 7, 2008
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0801017105