Hammann, Christian ; Martinez, E. ; Moosbauer, J. ; Hormes, R. ; Tabler, M. (1999)
Length variation of helix III in a hammerhead ribozyme and its influence on cleavage activity.
In: Antisense & nucleic acid drug development, 9 (1)
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
The previously described HIV-1 directed hammerhead ribozyme 2as-Rz12 can form with its target RNA 2s helices I and III of 128 and 278 base pairs (bp). A series of derivatives was made in which helix III was truncated to 8, 5, 4, 3, and 2 nucleotides (nt). These asymmetric hammerhead ribozymes were tested for in vitro cleavage and for inhibition of HIV-1 replication in human cells. Truncation of helix III to 8 bp did not affect the in vitro cleavage potential of the parental catalytic antisense RNA 2as-Rz12. Further truncation of helix III led to decreased cleavage rates, with no measurable cleavage activity for the 2 bp construct. All catalytically active constructs showed complex cleavage kinetics. Three kinetic subpopulations of ribozyme-substrate complexes could be discriminated that were cleaved with fast or slow rates or not at all. Gel purification of preformed ribozyme-substrate complexes led to a significant increase in cleavage rates. However, the complex cleavage pattern remained. In mammalian cells, the helix III-truncated constructs showed the same but no increased inhibitory effect of the comparable antisense RNA on HIV-1 replication.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 1999 |
| Autor(en): | Hammann, Christian ; Martinez, E. ; Moosbauer, J. ; Hormes, R. ; Tabler, M. |
| Art des Eintrags: | Bibliographie |
| Titel: | Length variation of helix III in a hammerhead ribozyme and its influence on cleavage activity. |
| Sprache: | Englisch |
| Publikationsjahr: | 1999 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Antisense & nucleic acid drug development |
| Jahrgang/Volume einer Zeitschrift: | 9 |
| (Heft-)Nummer: | 1 |
| Kurzbeschreibung (Abstract): | The previously described HIV-1 directed hammerhead ribozyme 2as-Rz12 can form with its target RNA 2s helices I and III of 128 and 278 base pairs (bp). A series of derivatives was made in which helix III was truncated to 8, 5, 4, 3, and 2 nucleotides (nt). These asymmetric hammerhead ribozymes were tested for in vitro cleavage and for inhibition of HIV-1 replication in human cells. Truncation of helix III to 8 bp did not affect the in vitro cleavage potential of the parental catalytic antisense RNA 2as-Rz12. Further truncation of helix III led to decreased cleavage rates, with no measurable cleavage activity for the 2 bp construct. All catalytically active constructs showed complex cleavage kinetics. Three kinetic subpopulations of ribozyme-substrate complexes could be discriminated that were cleaved with fast or slow rates or not at all. Gel purification of preformed ribozyme-substrate complexes led to a significant increase in cleavage rates. However, the complex cleavage pattern remained. In mammalian cells, the helix III-truncated constructs showed the same but no increased inhibitory effect of the comparable antisense RNA on HIV-1 replication. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie ?? fb10_mikrobiologie ?? 10 Fachbereich Biologie > Regulatorische RNAs und Ribozyme |
| Hinterlegungsdatum: | 29 Jul 2010 12:09 |
| Letzte Änderung: | 05 Mär 2013 09:34 |
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