Aptamer-Peptide Conjugate for Toxicity-Free Cellular Uptake
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for delivering L-RNA aptamers into cells are limited by poor cell penetration and require toxic transfection agents like LIPOFECTAMINE®, which restricts their biological applications.
Innovation Solution
An aptamer-peptide conjugate is developed, comprising an L-RNA aptamer linked to a cell-penetrating peptide (CPP) via a Cu(I)-catalyzed azide-alkyne cycloaddition reaction, facilitating cellular uptake without the need for transfection agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If L-RNA aptamers are delivered using traditional transfection agents like LIPOFECTAMINE®, then cellular uptake efficiency is improved, but cell toxicity increases
Solution Approach 1:
The patent introduces a cell-penetrating peptide (CPP) as an intermediary component that mediates the cellular uptake of L-RNA aptamers. The CPP is covalently linked to the L-RNA aptamer through a linker molecule, forming a conjugate that can efficiently enter cells without requiring toxic transfection agents. The CPP acts as a carrier that facilitates membrane penetration while the L-RNA aptamer maintains its binding function.
Solution Approach 2:
The patent creates a composite structure by chemically conjugating the L-RNA aptamer with the cell-penetrating peptide through a linker. This composite conjugate combines the cell-penetration capability of the CPP with the specific binding capability of the L-RNA aptamer, achieving both efficient cellular uptake and low toxicity.
2Reliability
If L-RNA aptamers are used without modification, then binding specificity to G-quadruplex structures is maintained, but cell penetration capacity remains poor
Solution Approach 1:
The patent merges two distinct functional components into a single conjugate molecule: the cell-penetrating peptide (CPP) for membrane translocation and the L-RNA aptamer for specific G-quadruplex binding. The linker molecule connects these two components, allowing the conjugate to perform both functions sequentially - first penetrating the cell membrane, then binding to the target G-quadruplex structure inside the cell.
Solution Approach 2:
The patent applies local quality modification by adding the CPP and linker only to specific regions of the L-RNA aptamer structure. The L-RNA aptamer portion retains its native sequence and secondary structure necessary for G-quadruplex binding, while the attached CPP provides the cell-penetration function. This localized modification ensures that the binding specificity is preserved while gaining penetration capability.
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 conjugate efficiently translocates into cells and specifically binds to G-quadruplex structures, regulating gene activity and expression, achieving comparable uptake efficiency to traditional transfection methods without the use of toxic agents.
Implementation Method 1
the L-RNA aptamer is linked to the penetrating moiety via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction occurred between the alkyne group of the L-RNA aptamer and the azide group of the modified amino acid residue
Data Source
AI summary
Disclosed herein is an aptamer-peptide conjugate comprising a penetrating moiety and an L-form ribonucleic acid (L-RNA) aptamer linked thereto. According to some embodiments of the present disclosure, the L-RNA aptamer has an alkyne group linked to its 5′ end, and the penetrating moiety comprises a cell-penetrating peptide (CPP), a modified amino acid residue, and a first linker linking the modified amino acid residue to the CPP, in which the side chain of the modified amino acid residue has an azide group, so that the L-RNA aptamer is linked to the penetrating moiety via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction.