Virion-associated cofactor high-mobility group DNA-binding protein-1 facilitates transposition from the herpes simplex virus/Sleeping Beauty amplicon vector platform.
The development of the integration-competent, herpes simplex virus/Sleeping Beauty (HSV/SB) amplicon vector platform has created a means to efficiently and stably deliver therapeutic transcription units (termed "transgenons") to neurons within the mammalian brain. Furthermore, an investigation into the transposition capacity of the HSV/SB vector system revealed that the amplicon genome provides an optimal substrate for the transposition of transgenons at least 12 kb in length [de Silva, S., Mastrangelo, M.A., Lotta, L.T., Jr., Burris, C.A., Federoff, H.J., and Bowers, W.J. ( 2010 ). Gene Ther. 17, 424-431]. These results prompted an investigation into the factors that may contribute toward efficient transposition from the HSV/SB amplicon. One of the cellular cofactors known to play a key role during SB-mediated transposition is the high-mobility group DNA-binding protein-1 (HMGB1). Our present investigation into the role of HMGB1 during amplicon-based transposition revealed that transposition is not strictly dependent on the presence of cellular HMGB1, contrary to what had been previously demonstrated with plasmid-based SB transposition. We have shown for the first time that during amplicon preparation, biologically active HMGB1 derived from the packaging cell line is copackaged into amplicon vector particles. As a result, HSV/SB amplicon virions arrive prearmed with HMGB1 protein at levels sufficient for facilitating SB-mediated transposition in the transduced mammalian cell.
de Silva
Suresh
S
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.
Lotta
Louis T
LT
Burris
Clark A
CA
Bowers
William J
WJ
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004251:DNA Transposable Elements; D018014:Gene Transfer Techniques; D005809:Genes, Regulator; D005822:Genetic Vectors; D024243:HMGB1 Protein / Q000235:genetics*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D018139:Simplexvirus / Q000235:genetics*; D014162:Transfection; D019895:Transposases / Q000235:genetics*; D014771:Virion / Q000235:genetics*; D016662:Virus Integration
D004251:DNA Transposable Elements; D024243:HMGB1 Protein; D019895:Transposases; C494471:sleeping beauty transposase, human