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

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...

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Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2016-01, Vol.80 (1), p.114-120
Main Authors: Ichida, Yu, Utsunomiya, Yuko, Tomikawa, Junko, Nakabayashi, Kazuhiko, Sato, Toshinori, Onodera, Masafumi
Format: Article
Language:English
Subjects:
Animals
Binding Sites
Biological Assay
Cell Line, Tumor
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Gene Expression
Gene Silencing
Genes, Reporter
Genetic Vectors - chemistry
Genetic Vectors - metabolism
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
HEK293 Cells
Humans
K562 Cells
Mice
Moloney murine leukemia virus - chemistry
Moloney murine leukemia virus - genetics
Moloney murine leukemia virus - metabolism
Promoter Regions, Genetic
Protein Binding
retroviral vector
Stem Cells - metabolism
Stem Cells - pathology
Time Factors
zinc finger protein 809 (ZFP809)
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Summary: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. Time-course monitoring of gene silencing
ISSN:0916-8451
1347-6947
DOI:10.1080/09168451.2015.1072461