Long time-course monitoring of ZFP809-mediated gene silencing in transgene expression driven by promoters containing MLV-derived PBS

Yu Ichida, Yuko Utsunomiya, Junko Tomikawa, Kazuhiko Nakabayashi, Toshinori Sato, Masafumi Onodera

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Expression of Moloney murine leukemia virus (MoMLV)-typed retroviral vectors is strictly suppressed in immature cells such as embryonic stem cells. The phenomenon known as gene silencing is primed by the sequence-specific binding of the zinc finger protein 809 (ZFP809) to the primer-binding site of the vectors. However, it has yet to be determined whether the ZFP809-mediated gene silencing is maintained over a long period. In this study, we established an experimental system that can monitor gene silencing during a long-term cell culture using flow cytometry technology combined with fluorescent reporters for the expression of ZFP809 and the transgene expression driven by the promoters of interest. Time-course analysis using our system revealed that ZFP809 maintains gene silencing effect even at a longtime period. Furthermore, our system was useful for the monitoring of ZFP809-mediated gene silencing regardless of the types of vectors and cell lines.

Original languageEnglish
Pages (from-to)114-120
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume80
Issue number1
DOIs
Publication statusPublished - 2016

Keywords

  • Gene silencing
  • Retroviral vector
  • Zinc finger protein 809 (ZFP809)

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry

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