Cyclized L-RNA Aptamers for Nuclease Resistance
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Solution Overview
Problem
Existing L-RNA aptamers face challenges in achieving enhanced stability and target recognition due to limited modification approaches, making them susceptible to nuclease degradation and lacking conformational stability, binding affinity, and specificity.
Innovation Solution
Cyclization of L-RNA aptamers using intramolecular click chemistry with Cu(I)-catalyzed azide-alkyne cycloaddition, forming stable cyclized structures with 5′-alkyne and 3′-azide residues, enhancing stability and binding properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If L-RNA aptamers are used for target recognition, then binding affinity and specificity can be achieved, but the aptamers are susceptible to nuclease degradation and lack conformational stability
Solution Approach 1:
The patent applies parameter changes by modifying the chemical structure of the aptamer through cyclization. The linear L-RNA aptamer is transformed into a cyclic structure by forming a covalent bond between the 5' and 3' ends, which fundamentally changes the structural parameters and confers resistance to nuclease degradation while maintaining target binding capability
Solution Approach 2:
The patent creates a composite structure by combining the L-RNA aptamer sequence with a cyclic backbone formed through click chemistry. This composite cyclic structure integrates the target-recognition functionality of the aptamer with the enhanced stability of the cyclic framework, achieving both binding affinity and nuclease resistance
2Reliability
If linear L-RNA aptamers are synthesized, then target recognition capability is achieved, but stability and binding properties are limited
Solution Approach 1:
The patent transforms the linear configuration into a cyclic configuration, fundamentally changing the structural parameters of the aptamer. This cyclization process enhances conformational stability and improves binding properties by restricting the aptamer to its active folded structure, while preserving target recognition capability
Solution Approach 2:
The patent introduces curvature by converting the linear aptamer into a cyclic structure. This curved/closed configuration provides enhanced conformational stability and protects the aptamer from degradation, while maintaining the three-dimensional structure necessary for target recognition
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method results in L-RNA aptamers with improved resistance to nuclease degradation and enhanced target recognition, demonstrating strong interaction with G-quadruplex structures and effective inhibition of telomerase activity.
Implementation Method 1
ligating the L-RNA aptamer with a 3′ azide residue and a 5′ terminal alkyne group using intramolecular click chemistry reaction under the catalysis of Cu(I) to form a cyclized aptamer
Implementation Method 2
under the catalysis of Cu(I) to form a cyclized aptamer
Implementation Method 3
The present disclosure generally relates to aptamers and more particularly methods for cyclization of L-RNA aptamers to provide molecules having resistance to nuclease degradation
Implementation Method 4
enhancing stability and binding properties... enhanced target recognition
Data Source
AI summary
A method of cyclizing an L-RNA aptamer by modifying the aptamer with a 3′ azide and a 5′ alkyne group and using click chemistry reaction-based method. The cyclized L-RNA aptamers have improved binding properties and favour more in vitro/cell applications. Also disclosed is an L-oligonucleotide aptamer having linked ends.