In vitro selection of DNAzymes selective for monovalent metal ions and their development as sensors in living cells and organisms
type: CreativeWork
description: Beyond the function of carrying genetic information, the recognition function (DNA aptamers), the structural function (DNA origami and DNA nanodevices), and most interestingly, the catalytic function (deoxyribozymes or DNAzymes) of DNAs have been explored since 1990s, which attract widespread interest. DNAzymes, fully biocompatible but not naturally identified, commonly use metal ions as cofactors for catalysis, and some of them show exceptional selectivity for target metal ions over other metal ions, granting DNAzymes ideal advantages in metal ion sensing. To obtain such ion selective DNAzymes, in vitro selection methodology is exclusively used, and has resulted in a spectrum of RNA-cleaving DNAzymes selective for different types of metal ions, which have been leveraged for the development of environmental or live cell metal ion sensors. Potassium ion (K+) is the most abundant metal ion in all living cells and plays important biological roles. Hence, methods to detect K+ in living systems with high selectivity are invaluable to understand the precise mechanisms of K+ in physiological events. As the first step of live detection of K+, DNAzymes that are selective for K+ were obtained through in vitro selection and are being validated and truncated, which is recorded in the Chapter 2. The first K+-selective RNA-cleaving DNAzymes has been obtained from an in vitro selection, showing more than 16% cleavage yield by 20 hours under selection conditions with over 30-fold selectivity over Na+ and over 100-fold selectivity over other metal ions, and without restrictions by anion types. Since the proposal of “in vitro evolution” in 1989 and three independent successes in 1990 by the Joyce, Szostak, and Gold labs, in vitro selection (or SELEX) has been broadly used in obtaining and improving the performance of functional nucleic acids. To improve the effectiveness of selection, evolutionary stringency or pressure are usually applied during the selection process. However, how t…
abstract: Beyond the function of carrying genetic information, the recognition function (DNA aptamers), the structural function (DNA origami and DNA nanodevices), and most interestingly, the catalytic function (deoxyribozymes or DNAzymes) of DNAs have been explored since 1990s, which attract widespread interest. DNAzymes, fully biocompatible but not naturally identified, commonly use metal ions as cofactors for catalysis, and some of them show exceptional selectivity for target metal ions over other metal ions, granting DNAzymes ideal advantages in metal ion sensing. To obtain such ion selective DNAzymes, in vitro selection methodology is exclusively used, and has resulted in a spectrum of RNA-cleaving DNAzymes selective for different types of metal ions, which have been leveraged for the development of environmental or live cell metal ion sensors. Potassium ion (K+) is the most abundant metal ion in all living cells and plays important biological roles. Hence, methods to detect K+ in living systems with high selectivity are invaluable to understand the precise mechanisms of K+ in physiological events. As the first step of live detection of K+, DNAzymes that are selective for K+ were obtained through in vitro selection and are being validated and truncated, which is recorded in the Chapter 2. The first K+-selective RNA-cleaving DNAzymes has been obtained from an in vitro selection, showing more than 16% cleavage yield by 20 hours under selection conditions with over 30-fold selectivity over Na+ and over 100-fold selectivity over other metal ions, and without restrictions by anion types. Since the proposal of “in vitro evolution” in 1989 and three independent successes in 1990 by the Joyce, Szostak, and Gold labs, in vitro selection (or SELEX) has been broadly used in obtaining and improving the performance of functional nucleic acids. To improve the effectiveness of selection, evolutionary stringency or pressure are usually applied during the selection process. However, how t…
author: Yang, Zhenglin
language: en
keywords: Deoxyribozyme, DNAzyme, in vitro selection, metal ion, lithium, potassium, zinc, magnesium, DNA handling, PCR bias, GC rich sequence PCR, sensor, live imaging, in vivo imaging, metal ion profiling, fluorescence protein sensor, genetically encoded sensor
date published: 2022-01-12T22:54:58Z
date modified: 2026-07-22T22:24:53Z
provider: University of Illinois - Urbana-Champaign
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Illinois at Urbana-Champaign

PropertyValue
property ID: Handle
value: http://hdl.handle.net/2142/113284