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

Fingerprint

Zinc Fingers
Gene Silencing
Transgenes
Zinc
Genes
Monitoring
Proteins
Moloney murine leukemia virus
Flow cytometry
Embryonic Stem Cells
Stem cells
Viruses
Cell culture
Flow Cytometry
Cell Culture Techniques
Binding Sites
Cells
Technology
Cell Line

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

Cite this

Long time-course monitoring of ZFP809-mediated gene silencing in transgene expression driven by promoters containing MLV-derived PBS. / Ichida, Yu; Utsunomiya, Yuko; Tomikawa, Junko; Nakabayashi, Kazuhiko; Sato, Toshinori; Onodera, Masafumi.

In: Bioscience, Biotechnology and Biochemistry, Vol. 80, No. 1, 2016, p. 114-120.

Research output: Contribution to journalArticle

Ichida, Yu ; Utsunomiya, Yuko ; Tomikawa, Junko ; Nakabayashi, Kazuhiko ; Sato, Toshinori ; Onodera, Masafumi. / Long time-course monitoring of ZFP809-mediated gene silencing in transgene expression driven by promoters containing MLV-derived PBS. In: Bioscience, Biotechnology and Biochemistry. 2016 ; Vol. 80, No. 1. pp. 114-120.
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