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Efficient Silencing of Gene Expression by an ASON-Bulge-DNAzyme Complex

文献类型： 外文期刊

作者： Yi, Jianzhong 1 ; Liu, Chengqian 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Inst Anim Husb Vet Sci, Shanghai, Peoples R China

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2011 年 6 卷 4 期

页码：

收录情况： SCI

摘要： Background: DNAzymes are DNA molecules that can directly cleave cognate mRNA, and have been developed to silence gene expression for research and clinical purposes. The advantage of DNAzymes over ribozymes is that they are inexpensive to produce and exhibit good stability. The "10-23 DNA enzyme'' is composed of a catalytic domain of 15 deoxynucleotides, flanked by two substrate-recognition domains of approximately eight nucleotides in each direction, which provides the complementary sequence required for specific binding to RNA substrates. However, these eight nucleotides might not afford sufficient binding energy to hold the RNA substrate along with the DNAzyme, which would interfere with the efficiency of the DNAzyme or cause side effects, such as the cleavage of non-cognate mRNAs. Methodology: In this study, we inserted a nonpairing bulge at the 59 end of the "10-23 DNA enzyme'' to enhance its efficiency and specificity. Different sizes of bulges were inserted at different positions in the 59 end of the DNAzyme. The non-matching bulge will avoid strong binding between the DNAzyme and target mRNA, which may interfere with the efficiency of the DNAzyme. Conclusions: Our novel DNAzyme constructs could efficiently silence the expression of target genes, proving a powerful tool for gene silencing. The results showed that the six oligo bulge was the most effective when the six oligo bulge was 12-15 bp away from the core catalytic domain.

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