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OBJECTIVE To study inhibitory effect of antisense oligodeoxynucleotides (asODNs) on genomic HDV ribozyme activity. METHODS Plasmid pHDVrz277 used to transcribe a 112 nucleotides (nt) genomic HDV ribozyme constructed earlier in our laboratory. It had genomic and antigenomic HDV ribozyme sequences from 665-941nt. The inhibitory effect of five asODNs on genomic HDV ribozyme activity was studied to select the potential effective sequences in vitro. RESULTS The results showed that asODN, being complementary to 683-703nt (stem I and self-cleavage site), completely blocked the activity of a 112nt genomic HDV ribozyme transcription in vitro. The inhibition rate of asODNs against 756-770nt (SSrB) and 721-735nt (SSrA) were 69.36% and 48.64% respectively. However, two asODNs covered the 736-750nt (stem IV) and 704-720nt (SSrC) showed almost no inhibitory effect on genomic HDV ribozyme. Fine sequence specificity testing at region 683-703nt revealed that asODN as small as 13 nt flanking the self-cleavage site (683-695nt) retained a high inhibition rate (> 90%). CONCLUSIONS The results suggested that stem I and SSrB may play an important role in genomic HDV ribozyme and asODNs can block genomic HDV ribozyme activity almost completely.
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