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ID 34740
file
creator
Shiozawa, Seiji
Okada, Yohei
Tomioka, Ikuo
Maeda, Takuji
Kanda, Akifumi
Shinohara, Haruka
Suemizu, Hiroshi
Okano, James Hirotaka
Sasaki, Erika
Okano, Hideyuki
NDC
Medical sciences
abstract
Nonhuman primate embryonic stem (ES) cells have vast promise for preclinical studies. Genetic modification in nonhuman primate ES cells is an essential technique for maximizing the potential of these cells. The common marmoset (Callithrix jacchus), a nonhuman primate, is expected to be a useful transgenic model for preclinical studies. However, genetic modification in common marmoset ES (cmES) cells has not yet been adequately developed. To establish efficient and stable genetic modifications in cmES cells, we inserted the enhanced green fluorescent protein (EGFP) gene with heterotypic lox sites into the beta-actin (ACTB) locus of the cmES cells using gene targeting. The resulting knock-in ES cells expressed EGFP ubiquitously under the control of the endogenous ACTB promoter. Using inserted heterotypic lox sites, we demonstrated Cre recombinase-mediated cassette exchange (RMCE) and successfully established a monomeric red fluorescent protein (mRFP) knock-in cmES cell line. Further, a herpes simplex virus-thymidine kinase (HSV-tk) knock-in cmES cell line was established using RMCE. The growth of tumor cells originating from the cell line was significantly suppressed by the administration of ganciclovir. Therefore, the HSV-tk/ganciclovir system is promising as a safeguard for stem cell therapy. The stable and ubiquitous expression of EGFP before RMCE enables cell fate to be tracked when the cells are transplanted into an animal. Moreover, the creation of a transgene acceptor locus for site-specific transgenesis will be a powerful tool, similar to the ROSA26 locus in mice.
journal title
Stem Cells and Development
volume
Volume 20
issue
Issue 9
start page
1587
end page
1600
date of issued
2011
publisher
Mary Ann Liebert, Inc.
issn
1547-3287
ncid
publisher doi
language
eng
nii type
Journal Article
HU type
Journal Articles
DCMI type
text
format
application/pdf
text version
publisher
rights
(c) 2011 by Mary Ann Liebert, Inc. All Rights are Reserved.
relation url
department
Natural Science Center for Basic Research and Development



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