Baculoviral Vector-Mediated Transient and Stable Transgene Expression in Human Embryonic Stem Cells

Zeng, Jieming; Du, Juan; Ying, Zhao; Palanisamy, Nallasivam; Wang, Shu

doi:10.1634/stemcells.2006-0616

Cited by 90 publications

(102 citation statements)

References 19 publications

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“…These observations contradicted with previous data reporting that the transgene cassette flanked by the ITR elements, with or without the rep gene, offered sustained expression in the rat brains (490 days), 24 in human neuronal cells 29 and in human embryonic stem cells (422 weeks). 30 The disparity probably arose from the differences in the intrinsic properties (for example, repertoire of cellular factors) or in the proliferation rates of the cell types. Should integration not occur efficiently, the ITR-flanking cassette was more likely to persist in the slow proliferating neuronal/stem cells for a longer period of time than in the rapidly proliferating HEK293 cells in which the transgene was diluted out.…”

Section: Discussionmentioning

confidence: 99%

“…The WPRE (woodchuck hepatitis virus post-transcriptional regulatory element), a cis-acting element that enhanced the baculovirus-mediated transgene expression in vertebrate cells, 46 was amplified from pFastBac1-CMV-EGFP-WPRE 30 (kindly provided by Professor Shu Wang of National University of Singapore) and inserted into pBac-hEA between the hEA gene and pA to yield pBac-hEA/w. The IR/ DR elements from pT2-BH were digested by SalI/NotI and cloned into pFastBacDpolhDp10 to yield pBac-T2.…”

Section: Preparation Of Recombinant Baculovirusesmentioning

confidence: 99%

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Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

et al. 2011

Gene Ther

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Antiangiogenesis is an appealing anticancer approach but requires continued presence of the antiangiogenic agents, which can be remedied by gene therapy. Baculovirus is an emerging gene delivery vector but only mediates transient expression (o7 days); thus, this study primarily aimed to develop a hybrid baculovirus for sustained antiangiogenic gene expression and cancer therapy. We first constructed plasmids featuring adeno-associated virus inverted terminal repeats (AAV ITRs), oriP/ Epstein-Barr virus-expressed nuclear antigen 1 (EBNA1) or Sleeping Beauty (SB) transposon and compared their efficacies in terms of persistent expression. In human embryonic kidney (HEK293) cells, AAV ITR failed to prolong the expression while oriP/EBNA1 moderately extended the expression to 35 days. In contrast, the SB system led to stable expression beyond 77 days even without antibiotic selection. Given this finding, we constructed a hybrid SB baculovirus expressing the SB transposase and harboring the transgene cassette flanked by inverted repeat/direct-repeat (IR/DR) elements recognizable by SB. The hybrid SB baculovirus efficiently transduced mammalian cells and mediated an expression duration longer than that by conventional baculoviruses, thanks to the transgene persistence and integration. The SB baculovirus (Bac-SB-T2hEA/w) expressing the antiangiogenic fusion protein comprising endostatin and angiostatin (hEA) also enabled prolonged hEA expression. With sustained hEA expression, Bac-SB-T2hEA/w repressed the angiogenesis in vivo, hindered the growth of two different tumors (prostate tumor allografts and human ovarian tumor xenografts) in mice and extended the life span of animals. These data altogether implicated the potential of the hybrid SB-baculovirus vector for prolonged hEA expression and for the treatment of multiple types of angiogenesis-dependent tumors.

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Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Recombinant Baculovirusesmentioning

confidence: 99%

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

et al. 2011

Gene Ther

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“…This insect virus is emerging as a potentially safe class of gene delivery vectors because of its inability to replicate or cause toxicity in mammalian cells. [19][20][21][22][23] Baculovirus-derived vectors are capable of transducing both dividing and non-dividing cells including human embryonic stem cells (hESCs) 24,25 and mesenchymal stem cells. 26,27 The great potential of NSCs in cancer therapy highlights the need for consistent and renewable sources for the collection or production of uniform human NSCs suitable for clinical applications.…”

Section: Introductionmentioning

confidence: 99%

RETRACTED ARTICLE: Targeted suicide gene therapy for glioma using human embryonic stem cell-derived neural stem cells genetically modified by baculoviral vectors

et al. 2011

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Tumor-tropic neural stem cells (NSCs) can be used in the Trojan horse approach as cellular vehicles for targeted delivery of therapeutic agents to distant tumor sites. To realize this cancer therapy potential, it is important to have a renewable source to generate large quantities of uniform human NSCs. Here, we reported that NSCs derived from HES1 human embryonic stem cell line were capable of migrating into intracranial glioma xenografts after systemic injection or after intracranial injection at a site distant from the tumor. To test whether the HES1-derived NSCs can be used for cancer gene therapy, we used a baculoviral vector to introduce the herpes simplex virus thymidine kinase suicide gene into the cells and demonstrated that baculovirusmediated transgene expression may last for at least 3 weeks in NSCs. After being injected into the cerebral hemisphere opposite the tumor site and in the presence of ganciclovir, NSCs expressing the suicide gene were able to inhibit the growth of human glioma xenografts and prolong survival of tumor-bearing mice. Our findings suggest that human embryonic stem cells could potentially serve as a clinically viable source for production of cellular vehicles suitable for targeted anticancer gene therapy.

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“…Baculovirus vectors are able to transduce a wide range of human cell lines, including primary cells such as bone marrow fibroblasts, neural cells, hepatocytes and mesenchymal stem cells. Zeng and coworkers (Zeng et al, 2007) have demonstrated their ability to transduce human embryonic stem cells without altering their characteristics. The hybrid vectors contained three promoters, one driving the expression of the complete baculovirus genome, the second expressing eGFP and the third (AAVp5) driving the expression of Rep78.…”

Section: Baculovirus/aavmentioning

confidence: 99%

New Vectors for Stable and Safe Gene Modification

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Cobo³

et al. 2011

Gene Therapy - Developments and Future Perspectives

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Baculoviral Vector-Mediated Transient and Stable Transgene Expression in Human Embryonic Stem Cells

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Cited by 90 publications

References 19 publications

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

RETRACTED ARTICLE: Targeted suicide gene therapy for glioma using human embryonic stem cell-derived neural stem cells genetically modified by baculoviral vectors

New Vectors for Stable and Safe Gene Modification

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