RRC ID |
38129
|
著者 |
Ichida Y, Utsunomiya Y, Tomikawa J, Nakabayashi K, Sato T, Onodera M.
|
タイトル |
Long time-course monitoring of ZFP809-mediated gene silencing in transgene expression driven by promoters containing MLV-derived PBS.
|
ジャーナル |
Biosci Biotechnol Biochem
|
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.
|
巻・号 |
80(1)
|
ページ |
114-20
|
公開日 |
2016-1-1
|
DOI |
10.1080/09168451.2015.1072461
|
PMID |
26252886
|
MeSH |
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
Stem Cells / metabolism*
Stem Cells / pathology
Time Factors
|
IF |
1.516
|
引用数 |
1
|
リソース情報 |
ヒト・動物細胞 |
F9 |