Activity of HDV ribozymes to trans-cleave HCV RNA.
World J Gastroenterol
; 8(4): 694-8, 2002 Aug.
Article
em En
| MEDLINE
| ID: mdl-12174380
ABSTRACT
AIM:
To explore whether HDV ribozymes have the ability to trans-cleave HCV RNA.METHODS:
Three HDV genomic ribozymes were designed and named RzC1, RzC2 and RzC3. The substrate RNA contained HCV RNA 5'-noncoding region and 5'-fragment of C region (5'-NCR-C). All the ribozymes and HCV RNA 5'-NCR-C were obtained by transcription in vitro from their DNA templates, and HCV RNA 5'-NCR-C was radiolabelled at its 5'-end. Under certain pH, temperature, appropriate concentration of Mg(2+) and deionized formamide, these ribozymes were respectively or simultaneously mixed with HCV RNA 5'-NCR-C and reacted for a certain time. The trans-cleavage reaction was stopped at different time points, and the products were separated with polyacrylamide gel electrophoresis (PAGE), displayed by autoradiography. Percentage of trans-cleaved products was measured to indicate the activity of HDV ribozymes.RESULTS:
RzC1 and RzC2 could trans-cleave 26 % and 21.8 % of HCV RNA 5'-NCR-C under our reaction conditions with 2.5 mol.L(-1) deionized formamide respectively. The percentage of HCV RNA 5'-NCR-C trans-cleaved by RzC1, RzC2 or combined usage of the three ribozymes increased with time, up to 24.9 %, 20.3 % and 37.3 % respectively at 90 min point. Almost no product from RzC3 was observed.CONCLUSION:
HDV ribozymes are able to trans-cleave specifically HCV RNA at certain sites under appropriate conditions, and combination of several ribozymes aiming at different target sites can trans-cleave the substrate more efficiently than using only one of them.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
RNA Viral
/
Vírus Delta da Hepatite
/
RNA Catalítico
/
Hepacivirus
Idioma:
En
Ano de publicação:
2002
Tipo de documento:
Article