Chromosomal transposition of PiggyBac in mouse embryonic stem cells
- PMID: 18579772
- PMCID: PMC2440425
- DOI: 10.1073/pnas.0801017105
Chromosomal transposition of PiggyBac in mouse embryonic stem cells
Abstract
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.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
![Fig. 1.](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/2453712/bin/zpq9990834940001.gif)
![Fig. 2.](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/2453712/bin/zpq9990834940002.gif)
![Fig. 3.](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/2453712/bin/zpq9990834940003.gif)
![Fig. 4.](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/2453712/bin/zpq9990834940004.gif)
![Fig. 5.](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/2453712/bin/zpq9990834940005.gif)
Similar articles
-
Chromosomal mobilization and reintegration of Sleeping Beauty and PiggyBac transposons.Genesis. 2009 Jun;47(6):404-8. doi: 10.1002/dvg.20508. Genesis. 2009. PMID: 19391106
-
Generation of an inducible and optimized piggyBac transposon system.Nucleic Acids Res. 2007;35(12):e87. doi: 10.1093/nar/gkm446. Epub 2007 Jun 18. Nucleic Acids Res. 2007. PMID: 17576687 Free PMC article.
-
A hyperactive piggyBac transposase for mammalian applications.Proc Natl Acad Sci U S A. 2011 Jan 25;108(4):1531-6. doi: 10.1073/pnas.1008322108. Epub 2011 Jan 4. Proc Natl Acad Sci U S A. 2011. PMID: 21205896 Free PMC article.
-
Germline mutagenesis mediated by Sleeping Beauty transposon system in mice.Genome Biol. 2007;8 Suppl 1(Suppl 1):S14. doi: 10.1186/gb-2007-8-s1-s14. Genome Biol. 2007. PMID: 18047691 Free PMC article. Review.
-
Sleeping Beauty Transposition.Microbiol Spectr. 2015 Apr;3(2):MDNA3-0042-2014. doi: 10.1128/microbiolspec.MDNA3-0042-2014. Microbiol Spectr. 2015. PMID: 26104705 Review.
Cited by
-
XIST dampens X chromosome activity in a SPEN-dependent manner during early human development.Nat Struct Mol Biol. 2024 Jun 4. doi: 10.1038/s41594-024-01325-3. Online ahead of print. Nat Struct Mol Biol. 2024. PMID: 38834912
-
Regional random mutagenesis driven by multiple sgRNAs and diverse on-target genome editing events to identify functionally important elements in non-coding regions.Open Biol. 2024 Apr;14(4):240007. doi: 10.1098/rsob.240007. Epub 2024 Apr 3. Open Biol. 2024. PMID: 38565160 Free PMC article.
-
Precise genetic engineering with piggyBac transposon in plants.Plant Biotechnol (Tokyo). 2023 Dec 25;40(4):255-262. doi: 10.5511/plantbiotechnology.23.0525a. Plant Biotechnol (Tokyo). 2023. PMID: 38434112 Free PMC article.
-
High-Level Production of scFv-Fc Antibody Using an Artificial Promoter System with Transcriptional Positive Feedback Loop of Transactivator in CHO Cells.Cells. 2023 Nov 16;12(22):2638. doi: 10.3390/cells12222638. Cells. 2023. PMID: 37998372 Free PMC article.
-
Response to: DNA transposon mechanisms and pathways of genotoxicity.Mol Ther. 2023 Oct 4;31(10):2816. doi: 10.1016/j.ymthe.2023.07.023. Epub 2023 Aug 14. Mol Ther. 2023. PMID: 37582361 No abstract available.
References
-
- Hayes F. Transposon-based strategies for microbial functional genomics and proteomics. Annu Rev Genet. 2003;37:3–29. - PubMed
-
- Spradling AC, Rubin GM. Transposition of cloned P elements into Drosophila germ line chromosomes. Science. 1982;218:341–347. - PubMed
-
- Bellen HJ, et al. P-element-mediated enhancer detection: A versatile method to study development in Drosophila. Genes Dev. 1989;3:1288–1300. - PubMed
-
- Thibault ST, et al. A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac. Nat Genet. 2004;36:283–287. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases