[Construction of the expression vectors of @entity44013 ribozymes and their intracellular inhibiting activity against XXXX RNA].

@entity1 @entity44013 @entity11620 @entity3531
OBJECTIVES: To investigate whether @entity44013 ribozymes can intracellularly inhibit @entity3531 RNA. METHODS: The @entity1 expression vectors, @entity11620 -RzC1, @entity11620 -RzC2 and @entity11620 -RzC3, containing ribozymes cDNA of RzC1, RzC2, and RzC3, were constructed targeting different @entity3531 -5' NCR-C RNA regions. Then the @entity3531 -positive fetal hepatocytes were transfected with these plasmids using liposome-mediated method. The inhibitory effects of @entity44013 ribozymes were evaluated by @entity3531 RNA quantitation in cultured cells and the supernatants. RESULTS: (1) All the three @entity44013 ribozymes were inserted into the expression vector. (2) Fetal hepatocytes were infected with @entity3531 proven by RT-PCR and fluorescent quantitative PCR and expressed @entity3531 NS3 and NS5 antigens by immunocytochemistry. (3) @entity44013 ribozymes inhibited the activity of the target @entity3531 RNA at expect positions in @entity3531 -positive hepatocytes. At 0.5 micromol/L, the inhibitory rate of @entity11620 -RzC1, @entity11620 -RzC2, and @entity11620 -RzC3 was 53.2%, 50.5 %, and 10.6% respectively. @entity11620 -RzC1 was used continuously for one week, showing the inhibitory rate of 60.7%, 64.2%, 68.4%, 71.9%, 78.8% and 83.1% on the 2nd, 3rd, 4th, 5th, 6th and 7th day. CONCLUSION: The inhibitory activity of @entity11620 -RzC1 (107-113nt) and @entity11620 -RzC2 (268-274nt) is greater than that of @entity11620 -RzC3 (345-351nt) in @entity3531 -positive hepatocytes.
