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 language | English |
|---|---|
| Pages (from-to) | 114-120 |
| Number of pages | 7 |
| Journal | Bioscience, Biotechnology and Biochemistry |
| Volume | 80 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2016 |
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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 journal › Article
}
TY - JOUR
T1 - Long time-course monitoring of ZFP809-mediated gene silencing in transgene expression driven by promoters containing MLV-derived PBS
AU - Ichida, Yu
AU - Utsunomiya, Yuko
AU - Tomikawa, Junko
AU - Nakabayashi, Kazuhiko
AU - Sato, Toshinori
AU - Onodera, Masafumi
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
KW - Gene silencing
KW - Retroviral vector
KW - Zinc finger protein 809 (ZFP809)
UR - http://www.scopus.com/inward/record.url?scp=84958982195&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84958982195&partnerID=8YFLogxK
U2 - 10.1080/09168451.2015.1072461
DO - 10.1080/09168451.2015.1072461
M3 - Article
C2 - 26252886
AN - SCOPUS:84958982195
VL - 80
SP - 114
EP - 120
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 1
ER -