Herpes Simplex Virus/Sleeping Beauty Vector-Based Embryonic Gene Transfer Using the HSB5 Mutant: Loss of Apparent Transposition Hyperactivity In Vivo
| Autor: | Zsuzsanna Izsvák, Suresh de Silva, Zoltán Ivics, William J. Bowers, Louis T. Lotta, Clark A. Burris, Michael A. Mastrangelo |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Transposable element
viruses Virus Integration Mutant Genetic Vectors Mutagenesis (molecular biology technique) Transposases Biology Viral vector Insertional mutagenesis Mice Genetics Animals Humans Simplexvirus Transgenes Molecular Biology Transposase Research Articles Genetic transfer Gene Transfer Techniques Genetic Therapy Amplicon Molecular biology Mice Inbred C57BL Mutagenesis Insertional HEK293 Cells DNA Transposable Elements Molecular Medicine |
| Popis: | The Sleeping Beauty (SB) transposon system has been successfully used as a gene delivery tool in nonviral and viral vector platforms. Since its initial reconstruction, a series of hyperactive mutants of SB have been generated. Questions remain as to whether the enhanced in vitro activities of these SB transposase mutants translate to the in vivo setting, and whether such increased integration efficiencies will ultimately compromise the safety profile of the transposon platform by raising the risk of genomic insertional mutagenesis. Herein, we compared the in vivo impact of a herpes simplex virus (HSV) amplicon-vectored "wild-type" SB transposase (SB10) and a "hyperactive" SB mutant (HSB5), codelivered in utero with the HSVT-βgeo transposable reporter amplicon vector to embryonic day 14.5 C57BL/6 mice. The SB10 and HSB5 transposases do not disparately affect the viability and development of injected mouse embryos. Quantitation of brain-resident βgeo expression on postnatal day 21 revealed that mice receiving HSB5 exhibited only a trending increase in transgene expression compared with the SB10-infused group, an outcome that did not mirror the marked enhancement of HSB5-mediated transposition observed in vitro. These findings indicate that in vivo application of hyperactive SB mutants, although not differentially genotoxic to the developing mouse embryo, does not necessarily provide a significant therapeutic advantage over the employment of a lesser active SB when delivered in the context of the HSV/SB amplicon platform. |
| Databáze: | OpenAIRE |
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