title: [Reconstruction of hepatitis D virus ribozymes and their ability to trans-cleave hepatitis C virus RNA].
abstract: OBJECTIVE
  To study whether reconstructed hepatitis D virus (HDV) ribozymes have the ability to trans-cleave hepatitis C virus (HCV) RNA.
  
  
  METHODS
  We reconstructed the stem IV and substrate-binding region of HDV genomic ribozymes, thus obtained three HDV ribozymes named RzC1, RzC2 and RzC3 expected to be able to trans-cleave HCV RNA at predicted sites. The substrate containing HCV RNA 5'-noncoding region (5'-NCR) and 5'-fragment of C region (HCV RNA 5'-NCR-C) was synthesized by transcription in vitro, then radiolabelled at its 5'-end. Under certain pH and appropriate concentration of Mg(2+) with or without deionized formamide, the ribozymes and their substrate were mixed at mol ratio of 100 : 1 and reacted for two hours. Trans-cleaved products were shown by denatured polyacrylamide gel electrophoresis and autoradiography, and the percentage of trans-cleaved substrate was calculated as the activity indicator of these ribozymes.
  
  
  RESULTS
  RzC1 and RzC2 were able to trans-cleave HCV RNA 5'-NCR-C site specifically, and their activity could be enhanced by certain concentration of deionized formamide. RzC3 could not trans-cleave the substrate.
  
  
  CONCLUSIONS
  Optimized HDV genomic ribozymes can trans-cleave HCV RNA.
authors: Institute of Infectious Diseases of PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Y
authors: Y
authors: Yu
authors: Q
authors: Q
authors: Mao
authors: Q
authors: Q
authors: Li
authors: Y
authors: Y
authors: Wang
authors: F
authors: F
authors: Min
mesh_terms: D020121:5' Untranslated Regions / Q000378:metabolism; D016174:Hepacivirus / Q000235:genetics*; D003698:Hepatitis Delta Virus / Q000201:enzymology*; D016337:RNA, Catalytic / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*
chemical_list: D020121:5' Untranslated Regions; D016337:RNA, Catalytic; D012367:RNA, Viral