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 i nto neuronal cells that is mediated by Cre/loxP recombination using re combinant 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. E xogenous Bcl-2 was clearly induced in PC12 cell lines carrying CALNLbc l-2 after infection with recombinant adenovirus producing recombinase Cre (AxCANCre). Dual infection with both AxCANCre and a recombinant ad enovirus bearing CALNLbcl-2 showed efficient delivery of exogenous Bcl -2 into a hybrid motoneuronal cell line and primary chicken spinal mot oneurons. The delivery of foreign Bcl-2 promoted survival of motoneuro ns in medium either containing or lacking trophic support. Thus, this strategy for delivery of exogenous Bcl-2 will be useful for studying n euronal death as well as for introducing foreign genes into postmitoti c neurons under the control of recombinase Cre.