Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved.
Efficient gene transfer and regulated transgene expression in primate embryonic stem (ES) cells are highly desirable for future applications of the cells. In the present study, we have examined using the nonintegrating Sendai virus (SeV) vector to introduce the green fluorescent protein (GFP) gene into non-human primate cynomolgus ES cells. The GFP gene was vigorously and stably expressed in the cynomolgus ES cells for a year. The cells were able to form fluorescent teratomas when transplanted into immunodeficient mice. They were also able to differentiate into fluorescent embryoid bodies, neurons, and mature blood cells. In addition, the GFP expression levels were reduced dose-dependently by the addition of an anti-RNA virus drug, ribavirin, to the culture. Thus, SeV vector will be a useful tool for efficient gene transfer into primate ES cells and the method of using antiviral drugs should allow further investigation for regulated SeV-mediated gene expression.
Center for Molecular Medicine, Jichi Medical School, Minamikawachi, Tochigi, Japan.
K
K
Sasaki
M
M
Inoue
H
H
Shibata
Y
Y
Ueda
S-I
SI
Muramatsu
T
T
Okada
M
M
Hasegawa
K
K
Ozawa
Y
Y
Hanazono
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006410:Hematopoiesis; D008252:Macaca fascicularis; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000473:pathology / Q000502:physiology*; D029082:Sendai virus / Q000235:genetics*; D013724:Teratoma; D014161:Transduction, Genetic / Q000379:methods
D049452:Green Fluorescent Proteins