DEVELOPMENT OF E-SRS (ENVIRONMENT-SENSING RESPONSE SYSTEM) AS A NOVEL METHOD TO DISTINGUISH GENETIC ENVIRONMENTS AND RESOLVE CLOSELY RELATED NUCLEIC ACID SEQUENCES
type: CreativeWork
description: Existing limitations of conventional Nucleic Acid (NA) detection prompted us to conduct a Proof-of-Concept of a novel NA sensing platform called environment-Sensing Response System (e-SRS), which could deliver a physical response upon sensing its NA Target Sequence (NTS). e-SRS is a NA sensing and response system with two components: 1) An RNA based sensor that changes conformation and activates upon binding specific NTS; 2) A Response System that is triggered by the activated sensor to initiate some physical response, such as emitting a fluorescent signal to indicate presence of the NTS, or other biomolecular actions like induction of gene expression. Its modular nature, whereby the sensor is separate from the Response System, allows e-SRS flexibility in adapting to different formats and applications. The ability to activate Response System after sensing enables the e-SRS sensor to serve as a signal transducer, which passes a signal of one form from the environment (e.g. presence of specific NA), to that of another form as produced by the Response System (e.g. activation of inducible expression system). A biomolecular signal transducer could function in more diverse ways than a biomolecular probe, and could be a powerful tool in research, diagnostics and therapy. After initial tests, an early design known as e-SIGE was unworkable, likely because the sensor¿s RNA folding was designed without computational secondary structure prediction. The RNA folding likely did not occur as intended. We redesigned the physical implementation to create the current e-SRS, adapting an existing allosteric ribozyme, the Maxizyme, as e-SRS sensor, employing computational secondary structure prediction. We were able to successfully test e-SRS in the test tube environment via Ribozyme Trans-cleavage Assays (RTA). Unsatisfied with RNA cleavage assays via Denaturing Polyacrylamide Gel Electrophoresis, we developed the Reporter Substrate (RS), which provided real time fluorescence reporting …
abstract: Existing limitations of conventional Nucleic Acid (NA) detection prompted us to conduct a Proof-of-Concept of a novel NA sensing platform called environment-Sensing Response System (e-SRS), which could deliver a physical response upon sensing its NA Target Sequence (NTS). e-SRS is a NA sensing and response system with two components: 1) An RNA based sensor that changes conformation and activates upon binding specific NTS; 2) A Response System that is triggered by the activated sensor to initiate some physical response, such as emitting a fluorescent signal to indicate presence of the NTS, or other biomolecular actions like induction of gene expression. Its modular nature, whereby the sensor is separate from the Response System, allows e-SRS flexibility in adapting to different formats and applications. The ability to activate Response System after sensing enables the e-SRS sensor to serve as a signal transducer, which passes a signal of one form from the environment (e.g. presence of specific NA), to that of another form as produced by the Response System (e.g. activation of inducible expression system). A biomolecular signal transducer could function in more diverse ways than a biomolecular probe, and could be a powerful tool in research, diagnostics and therapy. After initial tests, an early design known as e-SIGE was unworkable, likely because the sensor¿s RNA folding was designed without computational secondary structure prediction. The RNA folding likely did not occur as intended. We redesigned the physical implementation to create the current e-SRS, adapting an existing allosteric ribozyme, the Maxizyme, as e-SRS sensor, employing computational secondary structure prediction. We were able to successfully test e-SRS in the test tube environment via Ribozyme Trans-cleavage Assays (RTA). Unsatisfied with RNA cleavage assays via Denaturing Polyacrylamide Gel Electrophoresis, we developed the Reporter Substrate (RS), which provided real time fluorescence reporting …
author: LEONG SHIANG RONG (LIANG XIANGRONG)
keywords: Nucleic Acid detection, Maxizyme, Maxizyme design, RNA secondary structure, Dengue, Malaria
date published: 2009-12-28
date modified: 2026-07-24T03:31:51Z
provider: National University of Singapore
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: National University of Singapore