Characterization of pro-apoptotic gene Bak using hammerhead ribozymes

Bak gene encodes a 23kDa-protein homologue of apoptosis inhibitor Bcl-2. Unlike Bcl-2, Bak stimulates several apoptosis pathways, and by interacting with Bcl-2 or Bcl-XL, it counteracts the anti-cell death effect. Enforced expression of Bak induces rapid and extensive apoptosis of serum-deprived fibroblasts. In contrast, the mutation of the Bak gene was observed in several cancer cells, suggesting a deficiency of the Bak cellular function can cause a cancerous malignancy. Although many studies of Bak have been performed, the mechanisms by which Bak functions in mammalian cells remain unclear. In this study, we constructed a hammerhead ribozyme specifically targeted to the Bak mRNA for inhibition of the intracellular function of Bak. This Bak-ribozyme should be useful for a detailed functional analysis of Bak.