The combined use of viral transcriptional and post-transcriptional regulatory elements to improve baculovirus-mediated transient gene expression in human embryonic stem cells

被引:17
作者
Du, Juan [1 ,2 ]
Zeng, Jieming [1 ]
Zhao, Ying [1 ,2 ]
Boulaire, Jerome [1 ]
Wang, Shu [1 ,2 ]
机构
[1] Inst Bioengn & Nanotechnol, Singapore 138669, Singapore
[2] Natl Univ Singapore, Dept Biol Sci, Singapore 117543, Singapore
基金
英国医学研究理事会;
关键词
Baculovirus; Human embryonic stem cells; Transient transduction; Genetic modification; Transgene expression; SUSTAINED TRANSGENE EXPRESSION; NONINTEGRATING LENTIVIRAL VECTORS; GREEN FLUORESCENT PROTEIN; IN-VIVO; BETA PROMOTER; HUMAN GENOME; THERAPY; EFFICIENT; BRAIN; INTEGRATION;
D O I
10.1016/j.jbiosc.2009.06.017
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Transient gene expression is one possible approach to manipulate the signaling pathways that control the proliferation and differentiation of human embryonic stem (hES) cells. We tested in hES cells a range of baculoviral vectors with a human elongation factor-la promoter and various viral regulatory elements and observed the most dramatic augmenting effect on the transient expression when the promoter was used together with the human cytomegalovirus immediate-early gene enhancer and the woodchuck hepatitis virus post-transcriptional regulatory elements. This vector provided a 1.6-fold increase in the percentage of transduced cells (up to 72%) over a vector containing the elongation factor-1 alpha promoter alone. The effective baculoviral transduction of hES cells did not affect cell proliferation, expression of embryonic stem cell markers and teratoma formation. This new viral vector for temporary transgene expression might become a useful tool for developmental biology studies and biomedical applications of hES cells. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.
引用
收藏
页码:1 / 8
页数:8
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