CATALYZING ACTIVITY OF REPIRATORY SYNCITIAL VIRUS-SPECIFIC
DEOXYRIBOZYME (DNAZYME)
XD Zhao, XQ Yang & CA Zhao.
Children¡¯s Hospital, Chongqing University of
Medical Sciences, Chongqing 400014, China
Objective: RSV is the most
important respiratory pathogen of childhood, being responsible for about
half of all bronchiolitis and a quarter of all cases of pneumonia during
the first few months of life. The overall objective of this project is to
lay groundwork for treating patients with respiratory syncytial virus
infection by using a novel molecular biological tool, DNAzyme to target the
RSV viral RNA replication in epithelial cells of respiratory tract.
Methods: DNAzyme designed to cut
the RSV RNAs will be tested in both in votro cell free and cell culture
assays to determine the impact of such inhibition on the expression of RSV
viral protein, as well as the RSV replication, using ELISA, RT-PCR and
Northern Blot.
Results: Five of all synthesized 6
DNAzymes showed obvious catalyzing activity toward target RSV genomic RNA
and messenger RNA. A efficient transfer system was successfully established
to deliver the DNAzymes into target cells. Further study is being conducted
to demonstrate they have same effects in human respiratory epithelial cell
line, 9HTE.
Conclusion: It is considered that
DNAzyme will be a potential strategy in control of gene expression. With
designed specific binding arms, RSV-specific DNAzymes could be used to
treat RSV infection in children based on programmed study.