L-RNA Aptamer-ASO Conjugates for Selective APP RNA G-Quadruplex Targeting

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for selectively targeting G-quadruplex structures, particularly in the amyloid precursor protein (APP) gene, face challenges due to structural similarities among G-quadruplexes, limiting effective therapeutic interventions.

Innovation Solution

Development of an L-RNA aptamer-antisense oligonucleotide (ASO) conjugate that specifically recognizes and binds to the APP 3′-untranslated region RNA G-quadruplex structure, utilizing a ribonucleic acid sequence and deoxyribonucleic acid sequences to suppress APP expression and facilitate imaging.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional G4 targeting methods are used, then general G4 binding is achieved, but selective targeting of specific G4 structures (e.g., APP gene) is limited due to structural similarities among G-quadruplexes

Engineering Contradiction:

Improveselectivity of G4 targetingVSAvoidapplicability to different G4 structures

Core Design Contradiction:

Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The conjugate is divided into two functional segments: an L-RNA aptamer portion that recognizes the specific 3D structure of APP G4, and an ASO portion that provides sequence-specific binding to the APP gene. This segmentation allows each component to specialize in one aspect of target recognition, achieving high selectivity for the APP gene while maintaining the ability to bind G4 structures.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention creates a composite molecular conjugate combining L-RNA aptamer and ASO components linked by a stable bond. This composite structure integrates the structural recognition capability of the aptamer with the sequence-specific binding of the ASO, enabling simultaneous achievement of high selectivity for APP gene and effective G4 targeting.

Inventive Principle:

Principle #40Composite materials

2Measurement precision

If L-RNA aptamer-ASO conjugate is designed to specifically bind APP G4, then selectivity for APP gene is improved, but the complexity of the conjugate structure increases

Engineering Contradiction:

Improvespecificity of APP G4 bindingVSAvoidconjugate structure complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The L-RNA aptamer and ASO are merged into a single conjugate molecule through a stable bond linkage. This merging allows the conjugate to function as a unified entity that simultaneously provides structural recognition and sequence-specific binding, achieving high specificity for APP G4 while managing the overall structural complexity through integrated design.

Inventive Principle:

Principle #5Merging (Combining)

3Reliability

If the conjugate is used to suppress APP expression, then therapeutic effect is achieved, but the time and dose requirements for effective suppression must be optimized

Engineering Contradiction:

Improveeffectiveness of APP expression suppressionVSAvoidtime for expression suppression

Core Design Contradiction:

ReliabilityVSLoss of time

Solution Approach 1:

The conjugate continuously binds to APP G4 structures and suppresses expression through sustained interaction. The stable bond between conjugate components and the continuous binding action maintain effective suppression over time, reducing the need for high doses or extended treatment periods while ensuring reliable therapeutic effect.

Inventive Principle:

Principle #20Continuity of useful action

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 L-RNA aptamer-ASO conjugate effectively suppresses APP expression in a dose- and time-dependent manner, providing a tool for imaging and potentially treating neurodegenerative diseases like Alzheimer's disease by reducing APP protein levels.

Implementation Method 1

Guanine (G)-rich sequences of single-stranded DNA and RNA can fold into stable, intra-or intermolecular secondary structures called G-quadruplexes (dG4s and rG4s). Four guanines interact with each other by Hoogsteen-hydrogen bonds to form a planar structure, G-quartet. Stacking of two or more G-quartets, connected by loop nucleotides forms a G4 structure

Methodology Applied

Scientific EffectG-quadruplex structure formation:

Implementation Method 2

Four guanines interact with each other by Hoogsteen-hydrogen bonds to form a planar structure, G-quartet

Methodology Applied

Scientific EffectHoogsteen hydrogen bonding:

Implementation Method 3

the ASO comprises a deoxyribonucleic acid sequence selected from the group consisting of SEQ ID NOs: 2, 3 or 4

Methodology Applied

Scientific EffectAntisense oligonucleotide mechanism:

Data Source

PatentUS20250354149A1L-RNA aptamer-antisense oligonucleotide conjugates and uses thereof

Publication Date: 2025.11.20 CITY UNIVERSITY OF HONG KONG

AI summary

An L-form ribonucleic acid (L-RNA) aptamer-antisense oligonucleotide (ASO) conjugate comprise the L-RNA aptamer, which comprises a ribonucleic acid sequence. The ASO comprises a deoxyribonucleic acid sequence selected from a group. A method of imaging amyloid precursor protein (APP) rG4 in a cell comprising transfecting the cell with a messenger RNA of APP, permeating the cell, contacting the permeated cell with a cyanine3 (Cy3) labeled APP nucleic acid probe and the L-RNA aptamer-ASO conjugate, and subjecting the product to fluorescence microscopy analysis to produce an image of the APP rG4 in the cell.