Knockout serum replacement (KSR) has a suppressive effect on Sendai virus-mediated transduction of cynomolgus ES cells.
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.
Kishi
Yukiko
Y
Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, Tochigi, Japan.
Inoue
Makoto
M
Tanaka
Yujiro
Y
Shibata
Hiroaki
H
Masuda
Shigeo
S
Ikeda
Tamako
T
Hasegawa
Mamoru
M
Hanazono
Yutaka
Y
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008252:Macaca fascicularis; D051379:Mice; D008807:Mice, Inbred BALB C; D029082:Sendai virus / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D019076:Transgenes
D003470:Culture Media; D049452:Green Fluorescent Proteins