Proceedings of the National Academy of Sciences · 2013 · 231 citations · 26 references
Life SciencesGeneticsMolecular GeneticsGenetic EngineeringGenome EditingGenome EngineeringGenome Integration StrategiesSite-directed MutagenesisTransgenesis StrategiesTransgenic Organism DevelopmentTransgenic Organism ProductionTransposable ElementsInsertional MutagenesisPrecision Genome EngineeringTransgene IntegrationTransient TransgenesisTransposase FunctionTransposon ExcisionTransposon-mediated IntegrationTransposon-mediated Transgenesis
DNA transposons, such as piggyBac, excise and insert precisely, making them valuable for genome engineering and transient transgenesis like induced pluripotent stem cell production. Mutagenesis produced an Excision + Integration‑negative piggyBac transposase, likely targeting a DNA‑binding domain, that excises without reintegration.
Significance DNA transposons that translocate by excision from a donor site and insertion into a target site are often used for genome engineering by insertional mutagenesis and transgenesis. The piggyBac element is especially useful because it can excise precisely from an insertion site, restoring the site to its pretransposon state. Precise excision is particularly useful when transient transgenesis is needed, for example, in the transient introduction of transcription factors for induced pluripotent stem cell production. We have used mutagenesis to generate an Excision + Integration − transposase that allows piggyBac excision without potentially harmful reintegration. These mutations likely lie in a target DNA-binding domain.
26
Carrie Baker Brachmann, Adrian Davies, Gregory J. Cost et al. · Yeast · 1998 · 3.6K citations
piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells
Knut Woltjen, Iacovos P. Michael, Paria Mohseni et al. · Nature · 2009 · 1.8K citations · Full text
Efficient Transposition of the piggyBac (PB) Transposon in Mammalian Cells and Mice
Sheng Ding, Xiaohui Wu, Gang Li et al. · Cell · 2005 · 1K citations · Full text