| RRC ID |
38579
|
| 著者 |
Kakeda M, Nagata K, Osawa K, Matsuno H, Hiratsuka M, Sano A, Okazaki A, Shitara S, Nishikawa S, Masuya A, Hata T, Wako S, Osaki M, Kazuki Y, Oshimura M, Tomizuka K.
|
| タイトル |
A new chromosome 14-based human artificial chromosome (HAC) vector system for efficient transgene expression in human primary cells.
|
| ジャーナル |
Biochem Biophys Res Commun
|
| Abstract |
The use of non-integrating human artificial chromosomes (HACs) in gene therapy possibly allows for safe and reliable genetic modification of human cells without insertional mutagenesis and/or unexpected oncogene activations. Although we previously demonstrated that the HAC provides long-term therapeutic erythropoietin (EPO) production in normal human primary fibroblasts (hPFs), the expression level of EPO was too low to provide medical benefits for human therapy. Thus, the next challenge for the application of this system in therapeutic purposes is to improve the transgene expression on HACs. Here, we newly constructed chromosome 14-based HACs and examined the effects of the telomere and promoter regions on the expression level of the tansgene in hPFs. We showed that the use of natural telomere/sub-telomere and enhancers within the 5' untranslated region of the human ubiquitin C gene greatly increased (over 1000-fold) the EPO production in hPFs. Furthermore, we demonstrated the reprogramming of mouse embryonic fibroblasts by HAC-mediated introduction of four transcription factors, and established induced pluripotent stem cells with no trace of the HACs carrying multiple expression cassettes with large genome fragments. These results indicate that this HAC system could allow us to manipulate multiple transgenes efficiently in human primary cells, providing a promising tool not only for gene therapy but also for investigating genome functions in drug discoveries.
|
| 巻・号 |
415(3)
|
| ページ |
439-44
|
| 公開日 |
2011-11-25
|
| DOI |
10.1016/j.bbrc.2011.10.088
|
| PII |
S0006-291X(11)01900-0
|
| PMID |
22051050
|
| MeSH |
Animals
Cell Line
Cellular Reprogramming*
Chromosomes, Artificial, Human / genetics*
Chromosomes, Human, Pair 14 / genetics*
Enhancer Elements, Genetic / genetics
Erythropoietin / genetics
Fibroblasts
Gene Expression
Gene Transfer Techniques*
Genetic Therapy
Genetic Vectors*
Humans
Induced Pluripotent Stem Cells / metabolism
Mice
Promoter Regions, Genetic / genetics
Telomere / genetics
Transgenes*
|
| IF |
2.985
|
| 引用数 |
15
|
|
WOS 分野
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
|
