Steps toward targeted insertional mutagenesis with class II transposable elements

Sareina Chiung Yuan Wu, Kommineni J. Maragathavally, Craig J. Coates, Joseph M. Kaminski

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Insertional mutagenesis can be achieved by a variety of approaches, including both random and targeted methods. In contrast to chemical mutagenesis, insertional mutagens provide a molecular tag, thereby allowing rapid identification of the mutated genomic region. Integration into defined genomic locations has great utility for both gene insertion and mutagenesis. Our laboratories have explored targeted integration through the use of transposases coupled to defined DNA-binding domains. This technology holds great promise for targeted insertional mutagenesis by biasing integration events to regions recognized by the chosen DNA-binding domain. Herein, we provide a brief background on targeted transposon integration and detailed protocols for testing chimeric transposases in both mammalian cell culture and insect embryos.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages139-151
Number of pages13
Volume435
DOIs
StatePublished - Dec 1 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume435
ISSN (Print)10643745

Keywords

  • Cell culture
  • Chimeric transposases
  • DNA-binding domain
  • Gal4-UAS
  • Insertional mutagenesis
  • PiggyBac

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Wu, S. C. Y., Maragathavally, K. J., Coates, C. J., & Kaminski, J. M. (2008). Steps toward targeted insertional mutagenesis with class II transposable elements. In Methods in Molecular Biology (Vol. 435, pp. 139-151). (Methods in Molecular Biology; Vol. 435). https://doi.org/10.1007/978-1-59745-232-8-10