PITT

Pronuclear injection-based targeted transgenesis, a reliable transgene expression method in mice

Masato Ohtsuka, Hiromi Miura, Masahiro Sato, Minoru Kimura, Hidetoshi Inoko, Channabasavaiah B Gurumurthy

Research output: Contribution to journalReview article

18 Citations (Scopus)

Abstract

Transgenic (Tg) mice have been extensively used as valuable tools for analyses of gene function and have also served as models for many human diseases. Typically, a transgenic mouse is created by microinjection of DNA into pronuclei in which the DNA gets integrated at random locations in the genome. Frequently however, the random integration of multiple copies of a transgene results in transgene silencing, probably because of a positional effect and/or repeat-induced gene silencing. The transgene silencing issue has been overcome by single-copy transgene integration into a predetermined locus through ES cell-mediated transgenesis, despite it being expensive and more time-consuming compared with pronuclear injection (PI)-mediated transgenesis. Recently, several groups have reported novel approaches that employ PI for targeted transgenesis. They are based on site-specifc recombination catalyzed by a recombinase or an integrase or homologous recombination enhanced by a zinc-fnger nuclease via PI. These next-generation transgenesis methods, which we termed as PI-based Targeted Transgenesis (PITT), are more convenient and faster than ES cell-based transgenesis. Furthermore, the Tg mice generated by these newer methods contain a single-copy transgene and exhibit reliable expression of the transgene. The objective of this review is to present the recent progress in mouse targeted transgenesis.

Original languageEnglish (US)
Pages (from-to)489-502
Number of pages14
JournalExperimental Animals
Volume61
Issue number5
DOIs
StatePublished - Dec 1 2012

Fingerprint

Gene Transfer Techniques
Transgenes
transgenes
Genes
injection
Injections
mice
Integrases
Recombinases
Transgenic Mice
DNA
genetically modified organisms
Zinc
methodology
pronucleus
nucleases
Homologous Recombination
homologous recombination
Microinjections
Gene Silencing

Keywords

  • Mouse
  • PITT
  • Pronuclear injection
  • Targeted transgenesis
  • Transgene expression

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Biochemistry, Genetics and Molecular Biology(all)
  • veterinary(all)

Cite this

PITT : Pronuclear injection-based targeted transgenesis, a reliable transgene expression method in mice. / Ohtsuka, Masato; Miura, Hiromi; Sato, Masahiro; Kimura, Minoru; Inoko, Hidetoshi; Gurumurthy, Channabasavaiah B.

In: Experimental Animals, Vol. 61, No. 5, 01.12.2012, p. 489-502.

Research output: Contribution to journalReview article

Ohtsuka, Masato ; Miura, Hiromi ; Sato, Masahiro ; Kimura, Minoru ; Inoko, Hidetoshi ; Gurumurthy, Channabasavaiah B. / PITT : Pronuclear injection-based targeted transgenesis, a reliable transgene expression method in mice. In: Experimental Animals. 2012 ; Vol. 61, No. 5. pp. 489-502.
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