The combined use of viral transcriptional and post-transcriptional regulatory elements to improve baculovirus-mediated transient gene expression in human embryonic stem cells.
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-1alpha 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-1alpha 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.
Du
Juan
J
Institute of Bioengineering and Nanotechnology, Singapore, Singapore.
Zeng
Jieming
J
Zhao
Ying
Y
Boulaire
Jerome
J
Wang
Shu
S
D016367:Baculoviridae / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D005786:Gene Expression Regulation; D017781:Genes, Immediate-Early / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D020648:Peptide Elongation Factor 1 / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013724:Teratoma / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics
D020648:Peptide Elongation Factor 1