PMID- 19240695
OWN - NLM
STAT- MEDLINE
DCOM- 20090805
LR  - 20211020
IS  - 1525-0024 (Electronic)
IS  - 1525-0016 (Print)
IS  - 1525-0016 (Linking)
VI  - 17
IP  - 4
DP  - 2009 Apr
TI  - Development of a hybrid baculoviral vector for sustained transgene expression.
PG  - 658-66
LID - 10.1038/mt.2009.13 [doi]
AB  - Baculovirus is a promising gene delivery vector but its widespread application is 
      impeded as it only mediates transient transgene expression in mammalian cells. To 
      prolong the expression, we developed a dual baculovirus system whereby one 
      baculovirus expressed FLP recombinase while the other harbored an Frt-flanking 
      cassette encompassing the transgene and oriP/EBNA1 derived from Epstein-Barr 
      virus. After cotransduction of cells, the expressed FLP cleaved the Frt-flanking 
      cassette off the baculovirus genome and catalyzed circular episome formation, 
      then oriP/EBNA1 within the cassette enabled the self-replication of episomes. The 
      excision/recombination efficiency was remarkably enhanced by sodium butyrate, 
      reaching 75% in human embryonic kidney-293 (HEK293) cells, 85% in baby-hamster 
      kidney (BHK) cells, 77% in primary chondrocytes, and 48% in mesenchymal stem 
      cells (MSCs). The hybrid baculovirus substantially prolonged the transgene 
      expression to approximately 48 days without selection and >63 days with 
      selection, thanks to the maintenance of replicons and transgene transcription. In 
      contrast to the replicating episomes, the baculovirus genome was rapidly 
      degraded. Furthermore, an osteoinductive growth factor gene was efficiently 
      delivered into MSCs using this system, which not only prolonged the growth factor 
      expression but also potentiated the osteogenesis of MSCs. These data collectively 
      implicate the potential of this hybrid baculovirus system in gene therapy 
      applications necessitating sustained transgene expression.
FAU - Lo, Wen-Hsin
AU  - Lo WH
AD  - Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 
      Taiwan.
FAU - Hwang, Shiaw-Min
AU  - Hwang SM
FAU - Chuang, Ching-Kuang
AU  - Chuang CK
FAU - Chen, Chi-Yuan
AU  - Chen CY
FAU - Hu, Yu-Chen
AU  - Hu YC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090224
PL  - United States
TA  - Mol Ther
JT  - Molecular therapy : the journal of the American Society of Gene Therapy
JID - 100890581
RN  - 0 (Bone Morphogenetic Protein 2)
RN  - 0 (DNA Primers)
SB  - IM
MH  - Animals
MH  - Baculoviridae/*genetics
MH  - Base Sequence
MH  - Bone Morphogenetic Protein 2/genetics
MH  - Cell Line
MH  - Cricetinae
MH  - DNA Primers
MH  - *Genetic Vectors
MH  - Humans
MH  - Osteogenesis/genetics
MH  - Plasmids
MH  - Polymerase Chain Reaction
MH  - Recombination, Genetic
MH  - *Transgenes
PMC - PMC2835104
EDAT- 2009/02/26 09:00
MHDA- 2009/08/06 09:00
PMCR- 2010/04/01
CRDT- 2009/02/26 09:00
PHST- 2009/02/26 09:00 [entrez]
PHST- 2009/02/26 09:00 [pubmed]
PHST- 2009/08/06 09:00 [medline]
PHST- 2010/04/01 00:00 [pmc-release]
AID - S1525-0016(16)31559-3 [pii]
AID - 10.1038/mt.2009.13 [doi]
PST - ppublish
SO  - Mol Ther. 2009 Apr;17(4):658-66. doi: 10.1038/mt.2009.13. Epub 2009 Feb 24.