Targeting piggyBac transposon integrations in the human genome.

Galvan DL, Kettlun CS, Wilson MH
Methods Mol Biol. 2014 1114: 143-61

PMID: 24557901 · DOI:10.1007/978-1-62703-761-7_9

DNA based transposon systems offer a technology for active and efficient delivery of genes into human cells. An emerging field is directed at manipulating such systems to achieve site-directed integration as compared to un-targeted integration which occurs with native or unmodified transposon systems. The naturally active piggyBac transposon system is derived from insects but has been shown to be very efficient in gene-modifying human cells. Recent efforts have utilized the fusion of DNA binding domains to the piggyBac transposase enzyme with the goal of targeting integration to specific locations in the human genome. In this chapter, we describe methodology for engineering and characterizing chimeric piggyBac transposase enzymes, including experimental approaches for evaluating activity and targeting specificity in the human genome.

MeSH Terms (9)

DNA Transposable Elements Gene Order Gene Targeting Genetic Vectors Genome, Human HEK293 Cells Homologous Recombination Humans Mutagenesis, Insertional

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