Polysaccharide-dsRNA Complex for Cellular Uptake and Nuclease Resistance
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Solution Overview
Problem
Current RNA interference methods using double-stranded RNA face challenges with intracellular stability, cellular uptake, gene expression-inhibiting effect, and target specificity, and existing methods for transfecting double-stranded siRNA with β-1,3-glucan result in low transfection efficiency and reduced RNA interference effect.
Innovation Solution
A polysaccharide/double-stranded RNA complex is formed, where the double-stranded RNA has a sense strand and antisense strand complementary to a target gene sequence, with single-stranded polydeoxyadenine bound to the ends, and the polysaccharide forms a complex with polydeoxyadenine, enhancing cellular uptake and resistance to enzymatic degradation while maintaining RNA interference efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If double-stranded RNA is used for RNA interference, then gene expression-inhibiting effect is achieved, but intracellular stability and cellular uptake are insufficient
Solution Approach 1:
The patent combines double-stranded RNA with β-1,3-glucan to form a composite complex. The β-1,3-glucan component provides structural stability and protection against degradation, while the double-stranded RNA maintains its gene-silencing function. This composite structure resolves the contradiction by integrating the functional properties of both materials.
Solution Approach 2:
The patent introduces a carrier molecule (β-1,3-glucan or its derivatives) as an intermediary to deliver the double-stranded RNA into cells. This carrier mediates cellular uptake and protects the RNA from degradation, thereby improving intracellular stability without compromising the RNA interference effect.
2Reliability
If double-stranded RNA is used for RNA interference, then gene expression-inhibiting effect is achieved, but cellular uptake is insufficient
Solution Approach 1:
The patent uses β-1,3-glucan as a mediator to facilitate cellular uptake of double-stranded RNA. The carrier molecule interacts with cell surface receptors or membranes, enabling efficient internalization of the RNA complex while preserving its gene-silencing activity inside the cell.
Solution Approach 2:
The patent modifies the physical and chemical parameters of the delivery system by using β-1,3-glucan with specific molecular weight, charge, and structural properties. These parameter changes enhance cellular uptake efficiency through mechanisms such as endocytosis or membrane permeation, while maintaining the functional integrity of the double-stranded RNA.
3Stability of the object's composition
If terminal modification of double-stranded RNA is performed to enhance nuclease resistance, then nuclease resistance is improved, but RNA interference effect is greatly reduced
Solution Approach 1:
The patent separates the protective function from the functional RNA component. The β-1,3-glucan carrier provides nuclease resistance and protection, while the double-stranded RNA remains unmodified and retains its full RNA interference capability. This segmentation avoids the trade-off by placing the protective role on the carrier rather than modifying the RNA itself.
Solution Approach 2:
The patent uses β-1,3-glucan as an intermediary protective layer that shields the double-stranded RNA from nucleases without interfering with its gene-silencing function. The carrier absorbs the protective role, allowing the RNA to maintain its native structure and high RNA interference activity.
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 complex achieves improved cellular uptake and resistance to enzymatic degradation, maintaining the RNA interference effect, and can be used as a gene drug for treating diseases like cancer and AIDS, with the potential for additional functional molecule-mediated effects.
Implementation Method 1
the polysaccharide and the single-stranded polydeoxyadenine form a complex
Data Source
Figure 1A~1B
Figure 2~3
Figure 4
AI summary
An object of the present invention is to provide novel double-stranded RNA having an RNA interference effect, in which the cellular uptake and the resistance to enzymatic degradation are improved, without reducing the RNA interference effect. The cellular uptake and the resistance to enzymatic degradation of a complex of a polysaccharide having a β-1,3-glucan skeleton and double-stranded RNA can be significantly improved while maintaining the RNA interference effect, by fulfilling the following conditions (i) to (iii): (i) the double-stranded RNA has a sense strand consisting of a base sequence complementary to a target sequence in a target gene and an antisense strand containing a base sequence complementary to the sense strand, and the double-stranded RNA can inhibit expression of the target gene; (ii) the double-stranded RNA has a single-stranded polydeoxyadenine bound directly or via a linker to the end of at least one of the sense strand and the antisense strand; and (iii) the polysaccharide and the single-stranded polydeoxyadenine form a complex.