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Reselection Yielding a Smaller and More Active Silver-Specific DNAzyme.


ABSTRACT: Ag10c is a recently reported RNA-cleaving DNAzyme obtained from in vitro selection. Its cleavage activity selectively requires Ag+ ions, and thus it has been used as a sensor for Ag+ detection. However, the previous selection yielded very limited information regarding its sequence requirement, since only ?0.1% of the population in the final library were related to Ag10c and most other sequences were inactive. In this work, we performed a reselection by randomizing the 19 important nucleotides in Ag10c in such a way that a purine has an equal chance of being A or G, whereas a pyrimidine has an equal chance of being T or C. The round 3 library of the reselection was carefully analyzed and a statistic understanding of the relative importance of each nucleotide was obtained. At the same time, a more active mutant was identified, containing two mutated nucleotides. Further analysis indicated new base pairs leading to an enzyme with smaller catalytic loops but with ?200% activity of the original Ag10c, and also excellent selectivity for Ag+. Therefore, a more active mutant of Ag10c was obtained and further truncations were successfully performed, which might be better candidates for developing new biosensors for silver. A deeper biochemical understanding was also obtained using this reselection method.

SUBMITTER: Gu L 

PROVIDER: S-EPMC6643755 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature

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Reselection Yielding a Smaller and More Active Silver-Specific DNAzyme.

Gu Lide L   Saran Runjhun R   Yan Wanli W   Huang Po-Jung Jimmy PJ   Wang Shujun S   Lyu Mingsheng M   Liu Juewen J  

ACS omega 20181109 11


Ag10c is a recently reported RNA-cleaving DNAzyme obtained from in vitro selection. Its cleavage activity selectively requires Ag<sup>+</sup> ions, and thus it has been used as a sensor for Ag<sup>+</sup> detection. However, the previous selection yielded very limited information regarding its sequence requirement, since only ∼0.1% of the population in the final library were related to Ag10c and most other sequences were inactive. In this work, we performed a reselection by randomizing the 19 im  ...[more]

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