@article{Gu-2018-Reselection,
title = "Reselection Yielding a Smaller and More Active Silver-Specific DNAzyme",
author = "Gu, Lide and
Saran, Runjhun and
Yan, Wanli and
Huang, Po‐Jung Jimmy and
Wang, Shujun and
Lyu, Mingsheng and
Liu, Juewen",
journal = "ACS Omega, Volume 3, Issue 11",
volume = "3",
number = "11",
year = "2018",
publisher = "American Chemical Society (ACS)",
url = "https://gwf-uwaterloo.github.io/gwf-publications/G18-44001",
doi = "10.1021/acsomega.8b02039",
pages = "15174--15181",
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.",
}
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<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.</abstract>
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%0 Journal Article
%T Reselection Yielding a Smaller and More Active Silver-Specific DNAzyme
%A Gu, Lide
%A Saran, Runjhun
%A Yan, Wanli
%A Huang, Po‐Jung Jimmy
%A Wang, Shujun
%A Lyu, Mingsheng
%A Liu, Juewen
%J ACS Omega, Volume 3, Issue 11
%D 2018
%V 3
%N 11
%I American Chemical Society (ACS)
%F Gu-2018-Reselection
%X 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.
%R 10.1021/acsomega.8b02039
%U https://gwf-uwaterloo.github.io/gwf-publications/G18-44001
%U https://doi.org/10.1021/acsomega.8b02039
%P 15174-15181
Markdown (Informal)
[Reselection Yielding a Smaller and More Active Silver-Specific DNAzyme](https://gwf-uwaterloo.github.io/gwf-publications/G18-44001) (Gu et al., GWF 2018)
ACL
- Lide Gu, Runjhun Saran, Wanli Yan, Po‐Jung Jimmy Huang, Shujun Wang, Mingsheng Lyu, and Juewen Liu. 2018. Reselection Yielding a Smaller and More Active Silver-Specific DNAzyme. ACS Omega, Volume 3, Issue 11, 3(11):15174–15181.
