Knockout Serum Replacement (KSR) Has a Suppressive Effect on Sendai Virus-Mediated Transduction of Cynomolgus ES Cells
| Autor: | Yukiko Kishi, Shigeo Masuda, Mamoru Hasegawa, Yujiro Tanaka, Makoto Inoue, Tamako Ikeda, Yutaka Hanazono, Hiroaki Shibata |
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| Rok vydání: | 2008 |
| Předmět: |
viruses
Transgene Genetic Vectors Green Fluorescent Proteins Sendai virus Genome Green fluorescent protein Mice Transduction (genetics) Transduction Genetic Animals Transgenes Gene Cells Cultured Embryonic Stem Cells Mice Inbred BALB C biology respiratory system biology.organism_classification Embryonic stem cell Molecular biology Culture Media Macaca fascicularis Cattle Fetal bovine serum Developmental Biology Biotechnology |
| Zdroj: | Cloning and Stem Cells. 10:307-312 |
| ISSN: | 1557-7457 1536-2302 |
| DOI: | 10.1089/clo.2007.0080 |
| Popis: | Sendai virus (SeV) vectors can introduce foreign genes efficiently and stably into primate embryonic stem (ES) cells. For the application of these cells, the control of transgene expression is important. Cynomolgus ES cells transduced with a SeV vector expressing the green fluorescent protein (GFP) gene were propagated in Knockout serum replacement (KSR)-supplemented medium, used widely for the serum-free culture of ES cells, and growth and transgene expression were evaluated. The SeV vector-mediated GFP expression was suppressed in the KSR-supplemented medium, although it was stable in regular fetal bovine serum (FBS)-supplemented medium. Propagation in the KSR-supplemented medium eventually resulted in a complete suppression of GFP expression and eradication of the SeV genome. The inhibitory effect of KSR on the transduction was attributable to the positive selection of untransduced ES cells in addition to the removal of the SeV vector from transduced cells. KSR also reduced the efficiency of the transduction. SeV vector-mediated transgene expression in ES cells was suppressed in the KSR-supplemented medium. Although the suppression is limited in specified cells such as ES cells, these findings will help elucidate how to control transgene expression. |
| Databáze: | OpenAIRE |
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