MAO Jian-Ping 1)* ,WANG Quan-Hui 1) , HAN Hui-Ming 2) , YUAN Guo-Gang 1) , ZUO Gang 1) , SHAO Shi-He 2) , MAO Bing-Zhi 1) ( 1) Department of Immunology, Institute of Radiation Medicine, Beijing 100850, China; 2) College of Medicine,Beihua University, JiLin 132000, China)
10-23 DNAzyme is one of antisense oligo-deoxynucleotides (asODNs) which actively split mRNA molecules. The split activity was used for verifying the binding validity of asODNs to different targeting sites on mRNA. For 4 sites on the mRNA of green fluorescence protein (GFP), four asODNs and four 10-23 DNAzymes were designed in parallel. The control asODN had 2 mutation bases from asODN of the best site 2, and control 10-23 DNAzyme contained 2 mutation bases in the binding arms of the 10-23 DNAzyme to site 2.In vitro cleavage of GFP transcript by DNAzyme and asODNs dependent RNase H found that they have the same order in efficacy. After intracellular transfection of HeLa cells with 4 kinds of DNAzymes and 4 asODN,respectively,immuno_fluorescence microscopy examination indicated similar order in knockdown effects by reducing fluorescence intensity with most significant attenuation occurring for A4 and D4.The GFP knock down effectiveness by 10-23 DNAzyme were identical to those of respective asODNs as revealed by flow cytometry.These results suggested that 10-23 DNAzymes may be a novel tool for verifying the validity of target sites on gene mRNA.
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