Flexible Polycistronic RNAi Scaffold for Stoichiometric Delivery
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Solution Overview
Problem
Existing RNAi therapies face challenges in achieving proper molecular stoichiometries and specific ratios of siRNA molecules due to target site mutation, genetic variability, and off-target effects, particularly in treating complex diseases that require multiple molecular targets for effective silencing.
Innovation Solution
A reprogrammable polycistronic RNAi scaffold is developed, allowing administration of multiple RNAi sequences as a single RNA molecule with modular arms, enabling stoichiometric ratio control and simplifying production and characterization processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple siRNA molecules are used simultaneously to treat complex diseases, then therapeutic efficacy is improved, but manufacturing complexity and difficulty in achieving proper molecular stoichiometries increase
Solution Approach 1:
The patent merges multiple separate siRNA molecules into a single polycistronic scaffold molecule that contains multiple arms, each capable of producing different siRNA sequences. This combining approach maintains the therapeutic efficacy of multiple siRNAs while simplifying manufacturing to a single production process, thereby resolving the contradiction between therapeutic efficacy and manufacturing complexity.
Solution Approach 2:
The polycistronic scaffold serves as a universal platform that can produce multiple different siRNA sequences through its multiple arms, each with different seed sequences. This multi-functionality allows a single scaffold design to address multiple disease targets simultaneously, improving therapeutic efficacy while reducing the need for separate production processes for each siRNA.
2Reliability
If multiple siRNA molecules are administered to target different molecular targets, then treatment effectiveness is improved, but difficulty in achieving specific quantities and ratios of each siRNA increases
Solution Approach 1:
By merging multiple siRNA molecules into a single polycistronic scaffold with multiple arms, the patent ensures that each siRNA is produced in equal stoichiometric amounts during a single transcription process. This inherent equimolar production mechanism eliminates the difficulty of achieving specific quantities and ratios, as all siRNAs are synthesized simultaneously from the same scaffold template.
Solution Approach 2:
The scaffold is pre-designed with multiple arms that are positioned and configured to produce specific siRNA sequences in predetermined ratios. This preliminary arrangement of the arms and their corresponding seed sequences ensures that when the scaffold is processed, the correct stoichiometric ratios of different siRNAs are automatically generated, eliminating the need for post-production ratio adjustment.
3Ease of operation
If conventional drug delivery systems are used for RNAi administration, then ease of administration is improved, but ability to achieve proper molecular stoichiometries decreases
Solution Approach 1:
The patent combines multiple siRNA molecules into a single polycistronic scaffold that can be administered using conventional drug delivery systems. Because all siRNAs are produced simultaneously from the scaffold in equal amounts, the system maintains proper molecular stoichiometries while preserving the ease of administration through conventional delivery mechanisms, thereby resolving the contradiction between ease of operation and manufacturing precision.
Data Source
AI summary
Provided is a method for administering a single RNAi molecule or a combinatorial RNAi molecule via a polycistronic RNAi scaffold, where the polycistronic RNAi scaffold comprises one or more arms disposed on the polycistronic RNAi scaffold, wherein each of the one or more arms is an individual RNAi substrate, where at least one of the one or more arms comprise one or more of: a dicer independent loop sequence, a dicer independent loop-closing sequence, and a corresponding complement sequence. Further, at least one of the one or more arms may further comprise one or more of: an Ago2 cleavage sequence, a corresponding seed sequence, and a MID domain tight interaction.