RecQL1-Targeting siRNA Sequence With Serum-Stable RNAi Activity
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Solution Overview
Problem
Existing siRNAs targeting the RecQL1 helicase gene do not achieve superior effects in terms of RNAi activity against cancer cells while maintaining low toxicity to normal cells and stability in human serum.
Innovation Solution
Development of an siRNA with a novel target sequence comprising specific 2′-modified nucleosides and/or bridged nucleosides, along with modified internucleoside linkages, targeting the RecQL1 helicase gene, which enhances RNAi activity and stability in human serum.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional siRNA sequences targeting RecQL1 are used, then RNAi activity against cancer cells is achieved, but the effect is not superior in terms of RNAi activity, toxicity to normal cells, and stability in human serum
Solution Approach 1:
The patent applies parameter changes by modifying the chemical structure of the siRNA molecule. Specifically, it uses 2′-modified nucleosides (with 2′-O-methyl or 2′-fluoro groups) and bridged nucleosides at specific positions in the siRNA sequence. These chemical modifications change the physical and chemical parameters of the siRNA, thereby improving its RNAi activity, reducing toxicity to normal cells, and enhancing stability in human serum compared to conventional unmodified siRNAs.
Solution Approach 2:
The patent employs composite materials by combining different types of modified nucleosides within a single siRNA molecule. The siRNA contains a composite structure with 2′-modified nucleosides, bridged nucleosides, and modified internucleoside linkages all integrated into one molecule. This composite approach allows the siRNA to simultaneously achieve high RNAi activity, low toxicity, and high serum stability, which cannot be achieved with conventional homogeneous siRNA structures.
2Reliability
If conventional siRNA sequences are used, then cancer cell targeting is achieved, but stability in human serum is insufficient
Solution Approach 1:
The patent changes the chemical parameters of the siRNA by incorporating 2′-modified nucleosides and bridged nucleosides. These modifications alter the molecular structure to resist degradation by serum nucleases, thereby significantly improving stability and extending the half-life in human serum compared to conventional siRNAs.
Solution Approach 2:
The patent effectively replaces the short-living conventional siRNA with a modified version that has extended stability. The modified siRNA maintains its structural integrity and functional activity for a longer duration in serum, acting as a more durable therapeutic agent that persists longer in the biological system.
3Productivity
If existing target sequences are used, then RNAi activity is achieved, but no superior effect is obtained in terms of overall evaluation including efficacy
Solution Approach 1:
The patent optimizes the parameters of both the siRNA sequence and its chemical modifications to achieve superior efficacy. By carefully selecting the target sequence position (683-703 in RecQL1 mRNA) and combining it with specific nucleoside modifications at defined positions, the patent achieves enhanced RNAi activity and overall therapeutic effect compared to conventional sequences.
Solution Approach 2:
The patent applies local quality by introducing modifications at specific localized positions within the siRNA molecule rather than uniformly throughout. For example, 2′-modified nucleosides are placed at specific positions (such as positions 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22) and bridged nucleosides at other positions, creating localized regions of enhanced functionality that collectively improve overall efficacy while maintaining sequence specificity.
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 novel siRNA exhibits high therapeutic efficacy against tumor cells with reduced toxicity to normal cells, demonstrating improved RNAi activity and stability in human serum.
Implementation Method 1
an siRNA targeting a RecQL1 helicase gene, consisting of: a sense strand comprising the base sequence of SEQ ID NO: 1, and an antisense strand comprising the base sequence of SEQ ID NO: 2
Implementation Method 2
The present inventors have found as a result thereof an siRNA based on a novel target sequence, having properties far superior to those of conventional siRNAs in terms of RNAi activity against cancer cells, low toxicity to normal cells, and stability in human serum
Implementation Method 3
This siRNA has an activity of specifically inducing mitotic catastrophe and mitotic cell death to cancer cells
Data Source
AI summary
An siRNA targeting RecQL1 helicase gene, based on a target sequence, is provided having a superior effect as compared to conventional siRNAs. The siRNA targeting RecQL1 helicase gene includes a sense strand having the base sequence of SEQ ID NO: 1 and an antisense strand with the base sequence of SEQ ID NO: 2.