tailieunhanh - Báo cáo khoa học: An allosteric DNAzyme with dual RNA-cleaving and DNA-cleaving activities

A series of RNA-cleaving or DNA-cleaving DNAzymes have been obtained by in vitro selection. However, engineering an allosteric DNAzyme with dual RNA-cleaving and DNA-cleaving activities is very challenging. We used an in vitro-selected pistol-like (PL) DNAzyme as a DNA scaffold for designing a DNAzyme with dual catalytic activities. | ỊFEBS Journal An allosteric DNAzyme with dual RNA-cleaving and DNA-cleaving activities Dazhi Jiang Jiacui Xu Yongjie Sheng Yanhong Sun and Jin Zhang Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education Jilin University Changchun China Keywords activity allosteric DNAzyme grafting regulation Correspondence J. Zhang Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education Jilin University Changchun 130021 China Fax 86 431 88980440 Tel 86 431 88980440 E-mail zhangjin@ These authors contributed equally to this work Received 4 November 2009 revised 21 March 2010 accepted 1 April 2010 doi A series of RNA-cleaving or DNA-cleaving DNAzymes have been obtained by in vitro selection. However engineering an allosteric DNAzyme with dual RNA-cleaving and DNA-cleaving activities is very challenging. We used an in vitro-selected pistol-like PL DNAzyme as a DNA scaffold for designing a DNAzyme with dual catalytic activities. We prepared the 46-nucleotide DNAzyme with DNA-cleaving activity PL DNAzyme and then grafted the deoxyribonucleotide residues from an 8-17 variant DNAzyme into the region of stem-loop I and the catalytic core of the PL DNAzyme scaffold. This deoxyribonucleotide residue grafting resulted in a DNAzyme with dual RNA-cleaving and DNA-cleaving activities DRc DNAzyme . Drc DNAzyme has properties different from those of the original PL DNAzyme including DNA cleavage sites and the required metal ion concentration. Interestingly the RNA substrate and RNase A can act as effectors to mediate the DNA cleavage. Our results show that RNA-cleaving and DNA-cleaving activities simultaneously coexist in DRc DNAzyme and the DNA cleavage activity can be reversibly regulated by a conformational transition. Introduction DNAzymes are efficient biological catalysts that strengthen the catalytic power of nucleic acids 1 2 . To date a series of DNAzymes with RNA-cleaving or DNA-cleaving .

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