Cationic Morpholino Oligomers for Non-Invasive Gene Regulation
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Solution Overview
Problem
Morpholino-based antisense agents face challenges with poor solubility and complex synthesis procedures, leading to reduced antisense efficacy and the need for invasive microinjection methods for cell delivery, which limits their application in developmental biology and tissue culture studies.
Innovation Solution
Development of morpholino-based oligomers with a cationic backbone, incorporating guanidinium linkages and alkynyl/alkyl substitutions, which enhance solubility and transfection efficiency, allowing for cell penetration without injection and facilitating gene regulation studies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional morpholino-based antisense agents are used, then they can block gene expression, but they have poor solubility and require invasive microinjection methods for delivery
Solution Approach 1:
The patent modifies the chemical parameters of morpholino oligomers by introducing cationic groups (aminopropyl, guanidinium) at specific positions (N4, N6, N2 of purine bases; N3 of pyrimidine bases) to change the charge distribution and improve solubility, enabling non-invasive delivery while maintaining gene regulation efficacy
Solution Approach 2:
The patent creates composite morpholino structures by combining traditional morpholino backbone with cationic functional groups (aminopropyl, guanidinium) attached to nucleotide bases, forming a hybrid structure that integrates the gene-blocking capability of morpholinos with the solubility and cell-penetration properties of cationic groups
2Reliability
If traditional morpholino-based antisense agents are used, then they can inhibit gene function, but they have complex synthesis procedures and poor solubility
Solution Approach 1:
The patent divides the synthesis into modular stages: first synthesizing cationic morpholino monomers with specific functional groups (aminopropyl, guanidinium) at defined positions, then assembling these monomers into oligomers through phosphoramidite coupling, allowing systematic production of different sequences and modifications
Solution Approach 2:
The patent standardizes the chemical parameters of monomers by defining specific substitution patterns (N4, N6, N2, N3 positions) and protecting group strategies (acyl, benzoyl, trifluoroacetyl groups), creating a reproducible synthesis protocol that simplifies manufacturing while maintaining antisense efficacy
3Ease of operation
If cationic groups are introduced to improve solubility, then transfection efficiency increases, but the molecular structure becomes more complex
Solution Approach 1:
The patent applies cationic functional groups locally at specific positions (N4, N6, N2 of purines; N3 of pyrimidines) rather than uniformly across the entire oligomer, creating localized positive charge regions that enhance solubility and cell interaction while minimizing overall structural complexity
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 modified morpholino oligomers demonstrate improved solubility and transfection efficiency, enabling effective gene regulation and cellular penetration, thus overcoming the limitations of traditional morpholino agents, including reduced need for invasive delivery methods and increased specificity in gene regulation.
Implementation Method 1
The positive charges on the backbone give rise to cell membrane permeability through electrostatic attraction of the morpholinos to the negatively charged phosphate groups of the cell surface
Implementation Method 2
The binding, or hybridization, of antisense nucleic acid sequences to a specific mRNA target will, through a number of different mechanisms, interrupt normal cellular processing of the genetic message of a gene
Data Source
AI summary
Morpholino-based oligomers suitable as antisense agent comprising modifications of phosphorodiamidate backbone or modification with 5-substituted pyrimidines of morpholino compound that is soluble in culture medium and sufficient for cell penetration thereby eliminating the need for injecting into the cells. Monomers comprising the said oligomers and its method of manufacture, method of manufacture of the said oligomers and its dye, fluorophore, drug, biomolecule conjugate wherein the said oligomers find different end use but not limited to regulation of gene expression, tissue culture with improved transfection efficiency and related studies on cellular transfection.