Aptamer RNase Inhibitors for Heat-Stable RNA Sample Protection
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Solution Overview
Problem
Existing RNase inhibitors, particularly protein-based ones, are costly, inefficient, and unstable at high temperatures, leading to RNase contamination and degradation of RNA samples, which is a significant problem in RNA research and diagnostics.
Innovation Solution
Development of chemically synthesized single-stranded oligonucleotide aptamers that effectively inhibit both RNase A and RNase I, stable at high temperatures, and can be immobilized on matrices for enhanced efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If protein-based inhibitors are used to inhibit RNases, then RNase activity is inhibited, but the inhibitors are costly and unstable at high temperatures
Solution Approach 1:
The patent uses aptamers (nucleic acid molecules) as synthetic copies or alternatives to protein inhibitors. These aptamers are chemically synthesized oligonucleotides that bind to and inhibit RNases, providing a non-protein based solution that replicates the inhibitory function without the thermal instability and cost issues of protein-based inhibitors
Solution Approach 2:
The patent changes the chemical nature of the inhibitor from protein-based to nucleic acid-based (aptamers). This parameter change fundamentally alters the thermal stability profile, allowing the inhibitor to remain stable at high temperatures (up to 95°C) while maintaining RNase inhibition effectiveness
2Reliability
If large amounts of protein inhibitors are used to inhibit RNase activity, then RNase is effectively inhibited, but the cost increases significantly
Solution Approach 1:
The patent replaces expensive protein inhibitors with chemically synthesized oligonucleotide aptamers. These aptamers can be produced more cost-effectively through chemical synthesis rather than biological production and purification, reducing the overall cost while maintaining or improving inhibition effectiveness
Solution Approach 2:
The patent employs chemically synthesized oligonucleotide aptamers that are more cost-effective to produce than protein inhibitors. These synthetic molecules can be manufactured in quantities suitable for inhibition without the high costs associated with protein production, purification, and quality control
3Reliability
If protein inhibitors are used, then RNase inhibition is achieved, but the inhibitors require storage conditions that limit their shelf life
Solution Approach 1:
The patent changes the chemical composition from protein to nucleic acid (aptamer), which fundamentally improves storage stability. The aptamers can be stored at higher temperatures and for longer periods without degradation, extending their shelf life while maintaining inhibitory activity
Solution Approach 2:
The patent uses chemically synthesized oligonucleotide aptamers that can be formulated with various modifications to enhance stability. These synthetic molecules offer improved long-term stability profiles compared to protein inhibitors, maintaining activity after exposure to elevated temperatures
4Adaptability or versatility
If protein inhibitors are used to inhibit both RNase A and RNase I, then broad spectrum inhibition is achieved, but the inhibitors are expensive to manufacture and purify
Solution Approach 1:
The patent develops aptamers with broad-spectrum activity against multiple RNase types (including both RNase A and RNase I). This multi-functional capability is achieved through the selective binding properties of the aptamers, allowing a single inhibitor to target multiple RNase variants without requiring separate protein-based inhibitors for each type
Solution Approach 2:
The patent uses chemically synthesized oligonucleotide aptamers as alternatives to expensive protein inhibitors. These synthetic molecules can be produced more economically through chemical synthesis methods, reducing the manufacturing and purification costs while maintaining broad-spectrum inhibition capabilities
Data Source
AI summary
Compositions, methods and kits are provided that include an inhibitory oligonucleotide RNase inhibitor capable of inhibiting one or more types of RNase that coexist with biological samples or are introduced in the laboratory, thereby protecting RNA in the sample from degradation. More than one type of oligonucleotide RNase inhibitor may be combined in a mixture to inhibit a plurality of different RNases. Single oligonucleotides were identified to have inhibitory activity for a plurality of different RNases. The RNase oligonucleotide inhibitor may be immobilized on beads or other surface. It may be stored in a lyophilized form or in solution.