A novel strategy for introducing exogenous bcl-2 into neuronal cells: the Cre/loxP system-mediated activation of bcl-2 for preventing programmed cell death using recombinant adenoviruses.
We have established a novel strategy for introducing exogenous Bcl-2 into neuronal cells that is mediated by Cre/loxP recombination using recombinant adenoviral vectors. An on/off-switching cassette for Bcl-2 (CALNLbcl-2) was designed to express Bcl-2 by recombinase Cre-mediated excisional deletion of a spacer DNA flanked by a pair of loxP sites. Exogenous Bcl-2 was clearly induced in PC12 cell lines carrying CALNLbcl-2 after infection with recombinant adenovirus producing recombinase Cre (AxCANCre). Dual infection with both AxCANCre and a recombinant adenovirus bearing CALNLbcl-2 showed efficient delivery of exogenous Bcl-2 into a hybrid motoneuronal cell line and primary chicken spinal motoneurons. The delivery of foreign Bcl-2 promoted survival of motoneurons in medium either containing or lacking trophic support. Thus, this strategy for delivery of exogenous Bcl-2 will be useful for studying neuronal death as well as for introducing foreign genes into postmitotic neurons under the control of recombinase Cre.
Sato
N
N
Department of Neurobiology and Anatomy and Neuroscience Program, Wake Forest University School of Medicine, Winston-Salem, North Carolina, 27157, USA. nsato@bgsm.edu
Wang
S
S
Li
L
L
Okabe
K
K
Hashimoto
M
M
Yaginuma
H
H
Mikoshiba
K
K
Uchiyama
Y
Y
Uetsuki
T
T
Yoshikawa
K
K
Milligan
C E
CE
Oppenheim
R W
RW
D000256:Adenoviridae; D000257:Adenoviridae Infections / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002642:Chick Embryo; D004261:DNA Replication; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000502:physiology; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D007668:Kidney / Q000166:cytology; D009474:Neurons / Q000166:cytology* / Q000201:enzymology / Q000821:virology; D016716:PC12 Cells; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000502:physiology; D014161:Transduction, Genetic; D014764:Viral Proteins
D019253:Proto-Oncogene Proteins c-bcl-2; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases