Double-Stranded Antisense Nucleic Acid Complexes for Nuclear Delivery
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current nucleic acid drugs face challenges in delivering antisense oligonucleotides effectively into cell nuclei, particularly in achieving high specificity and efficiency for gene silencing and exon skipping applications.
Innovation Solution
Development of double-stranded nucleic acid complexes that include an antisense nucleic acid strand capable of inducing splice-switched variants of RNA, allowing for high specificity and efficiency in delivering antisense oligonucleotides into the nucleus, where they can modulate RNA processing and suppress target gene expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If single-stranded antisense oligonucleotides are used, then the structure is simple and easy to synthesize, but the delivery efficiency into cell nuclei is low
Solution Approach 1:
The patent combines a sense oligonucleotide and an antisense oligonucleotide to form a double-stranded complex. This merging of two separate components creates a structure that leverages cellular machinery for efficient nuclear delivery while maintaining the antisense functionality for gene silencing and exon skipping.
Solution Approach 2:
The sense oligonucleotide acts as an intermediary that facilitates nuclear entry. The double-stranded complex is recognized by cellular transport mechanisms that single-stranded oligonucleotides lack, enabling efficient delivery into the nucleus where the antisense strand can then exert its therapeutic effect.
2Productivity
If double-stranded nucleic acid complexes are used, then the delivery efficiency into cell nuclei is improved, but the structural complexity increases
Solution Approach 1:
The complex is segmented into two functional domains: the sense oligonucleotide that handles delivery and nuclear entry, and the antisense oligonucleotide that performs gene silencing and exon skipping. This segmentation allows each component to be optimized for its specific function while working together as a unified therapeutic agent.
Solution Approach 2:
The double-stranded complex serves multiple functions simultaneously: it acts as a delivery vehicle for nuclear entry, protects the antisense strand from degradation, and maintains the capability for specific target recognition and gene silencing. This multi-functionality justifies the increased structural complexity.
3Strength
If RNA oligonucleotides are used as ASO, then the binding affinity to target RNA is high, but the stability in vivo is reduced
Solution Approach 1:
Instead of using a single RNA strand that would be degraded, the patent creates a DNA-RNA hybrid complex where the DNA sense strand serves as a stable copy or template. This allows the RNA antisense strand to maintain high binding affinity while the DNA component provides enhanced in vivo stability and protection from nucleases.
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 double-stranded nucleic acid complexes demonstrate enhanced delivery and efficacy in suppressing target gene expression and altering RNA processing, offering potential therapeutic benefits for genetic diseases and other conditions by effectively reaching the nucleus and inducing desired exon skipping effects.
Implementation Method 1
an antisense oligonucleotide which has a sequence complementary to a transcription product of a target gene
Implementation Method 2
double-stranded nucleic acid complexes that include an antisense nucleic acid strand capable of inducing splice-switched variants of RNA, allowing for high specificity and efficiency in delivering antisense oligonucleotides into the nucleus
Implementation Method 3
a partial DNA-RNA hetero-duplex is formed. Since this structure is recognized by RNase H, and the mRNA of the target gene is thereby decomposed
Data Source
Figure 1
Figure 2
Figure 3
AI summary
Disclosed are double-stranded antisense nucleic acid complexes that can efficiently alter the processing of RNA in a cell via an antisense effect, and methods for using the same. One method comprises contacting with the cell a double-stranded nucleic acid complex comprising: a first nucleic acid strand annealed to a second nucleic acid strand, wherein: the first nucleic acid strand comprises (i) nucleotides independently selected from natural DNA nucleotides, modified DNA nucleotides, and nucleotide analogs, (ii) no regions that have 4 or more consecutive natural DNA nucleotides, (iii) the total number of natural DNA nucleotides, modified DNA nucleotides, and nucleotide analogs in the first nucleic acid strand is from 8 to 100, and (iv) the first nucleic acid strand is capable of hybridizing to RNA inside of the cell; and the second nucleic acid strand comprises nucleotides independently selected from natural RNA nucleotides, modified RNA nucleotides, and nucleotide analogs.