Designing and testing chimeric zinc finger transposases

Matthew H. Wilson, Alfred L. George*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

11 Scopus citations

Abstract

Transposons have been effectively utilized as non-viral gene delivery systems that are capable of promoting stable transgene expression in mammalian and human cells. Two specific transposon systems, Sleeping Beauty (SB) and piggyBac (PB), have been successfully modified by the addition of a zinc finger (ZF) DNA-binding domain as a strategy for directing transposon integration near ZF target sites in a host cell genome. Site-directed integration could improve transposon-mediated gene transfer by limiting positional effects on transgene integration, limiting genotoxicity, and improving the overall safety in gene therapy applications. In this chapter, we describe methodology for creating and characterizing functional chimeric ZF transposases and experimental approaches for testing their transpositional activity in human cells.

Original languageEnglish (US)
Title of host publicationEngineered Zinc Finger Proteins
Subtitle of host publicationMethods and Protocols
EditorsJoel Mackay, David Segal
Pages353-363
Number of pages11
DOIs
StatePublished - Dec 1 2010

Publication series

NameMethods in Molecular Biology
Volume649
ISSN (Print)1064-3745

Keywords

  • Transposon
  • human cells
  • piggyBac
  • site-directed integration
  • sleeping beauty
  • transposase
  • zinc finger

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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