Delta ribozyme benefits from a good stability in vitro that becomes outstanding in vivo.
The stability of a trans-acting delta ribozyme was studied under various conditions. Although in vitro (i.e., in the presence of protein extracts) this delta ribozyme appears to be only slightly more stable than a hammerhead ribozyme, in vivo (i.e., after cell transfection) it exhibits an outstanding stability that manifests itself in the calculated half-life of over 100 h regardless of the means of transfection. The P2 stem, which includes both the 5' and 3' ends, is shown to play a critical role in this stability. Direct mutagenesis of the most nuclease susceptible nucleotides failed to generate a more stable ribozyme that retained the same catalytic potential. Clearly, delta ribozyme appears to be well adapted to the human cell environment, and is therefore ideal for the development of a gene-inactivation system.
Lévesque
Dominique
D
Département de biochimie, Université de Sherbrooke, Quebec, Canada.
Choufani
Sanaa
S
Perreault
Jean-Pierre
JP
D001483:Base Sequence; D020134:Catalytic Domain; D002478:Cells, Cultured; D003698:Hepatitis Delta Virus / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D020871:RNA Stability; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship
D016337:RNA, Catalytic