Inhibition of Viral Multiplication in Acute and Chronic Stages of Infection by Ribozymes Targeted against the Polymerase Gene of Mouse Hepatitis Virus

Abstract

Two hammerhead ribozymes targeted against the polymerase gene of mouse hepatitis virus (MHV), which consisted of 22-nucleotide (nt) ribozyme core sequences and antisense sequences of different lengths, 243-nt (S-ribozyme) and 926-nt (L-ribozyme), were tested for their inhibitory effects on viral multiplication. Vectors that expressed the ribozymes were transfected into mouse DBT cells and several resulting cell lines constitutively expressing the ribozymes were selected and examined for intracellular MHV multiplication in acute and chronic stages of infection. The production of infectious progeny viral particles was significantly reduced in the transfected cell lines expressing either the S-ribozyme or L-ribozyme in acute infection. Although the in vitro cleavage process of the L-ribozyme was slower than that of the S-ribozyme, no difference was observed in inhibitory effects on MHV multiplication between S-and L-ribozymes in the transfected cells. In the transfected cells expressing L-ribozymes, production of viral particles was also inhibited in the chronic stage of MHV infection.