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.
Department of Neurobiology and Anatomy and Neuroscience Program, Wake Forest University School of Medicine, Winston-Salem, North Carolina, 27157, USA. nsato@bgsm.edu
N
N
Sato
S
S
Wang
L
L
Li
K
K
Okabe
M
M
Hashimoto
H
H
Yaginuma
K
K
Mikoshiba
Y
Y
Uchiyama
T
T
Uetsuki
K
K
Yoshikawa
C E
CE
Milligan
R W
RW
Oppenheim
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