DNA Construct With Multiple Promoters For OR Logic
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Solution Overview
Problem
Current methods for implementing OR logic in eukaryotic cells are complex, redundant, and inefficient, particularly for transcriptional and post-transcriptional levels, making it difficult to effectively analyze and manipulate heterogeneous cell populations for therapeutic and diagnostic applications.
Innovation Solution
A DNA construct with multiple promoters, each with its own transcriptional regulatory state, allows for the production of output RNA regardless of which promoter is activated, enabling OR gate-like logic operations in a single construct, reducing complexity and redundancy, and facilitating easier integration into viral vectors for therapeutic and diagnostic use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple distinct genetic constructs are used to implement OR logic, then the ability to target heterogeneous cell populations is improved, but the device complexity and payload size increase
Solution Approach 1:
The patent combines multiple promoter elements and their associated regulatory sequences into a single integrated genetic construct. This merging allows the construct to respond to multiple transcriptional inputs (TF1, TF2, etc.) through different promoters, achieving OR logic functionality while reducing the number of separate genetic constructs needed. The consolidated design decreases device complexity and payload size while maintaining the ability to target heterogeneous cell populations.
Solution Approach 2:
The single genetic construct is designed with multiple promoters that can each be activated by different transcription factors, making it universally applicable to detect and respond to various cell types and states. This multi-functional design allows one construct to perform what previously required multiple specialized constructs, thereby reducing complexity while enhancing adaptability to heterogeneous populations.
2Adaptability or versatility
If multiple distinct genetic constructs are used to implement OR logic, then the ability to target heterogeneous cell populations is improved, but the payload size increases preventing viral vector packaging
Solution Approach 1:
The patent merges multiple promoter-cassette elements into a single compact genetic construct that fits within viral vector packaging limits. By integrating multiple promoter elements and their regulatory sequences into one construct rather than using separate constructs, the total payload size is reduced while maintaining the ability to target heterogeneous cell populations through multiple transcriptional inputs.
3Adaptability or versatility
If two distinct genetic constructs are used for OR logic, then coverage of multiple cancer subtypes is improved, but output expression imbalance occurs
Solution Approach 1:
The patent combines multiple promoter elements under a single unified output sequence, ensuring that regardless of which promoter is activated (or if multiple are activated), the output expression level remains consistent. This merging eliminates the multi-valued logic problem where twice the output was obtained when both constructs were active, by normalizing the output through a shared downstream sequence.
4Adaptability or versatility
If redundant genetic information is included for OR logic implementation, then the ability to respond to multiple inputs is improved, but the construct becomes impractical for clinical translation
Solution Approach 1:
The patent integrates multiple promoter elements and their associated regulatory information into a single streamlined construct, eliminating redundancy while preserving the ability to respond to multiple transcriptional inputs. This consolidation makes the construct practical for clinical translation by reducing payload size and simplifying the genetic architecture, while still enabling responses to multiple cancer subtypes and cell states.
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 DNA construct achieves robust and simple OR gate-like logic operations, enabling effective classification and manipulation of different cell types and states, including cancer subtypes, with reduced payload size and improved scalability for therapeutic and diagnostic applications.
Implementation Method 1
each promoter comprises a transcription start site and/or is suitable for initiating transcription, wherein the initiation of transcription from the first promoter is enabled by a first transcriptional regulatory state and the initiation of transcription from each of said further promoter(s) is enabled by a respective further transcriptional regulatory state
Data Source
AI summary
The invention relates to the fields of biomolecular computing and/or synthetic biology. In particular, the invention relates to a DNA construct that comprises a first promoter, at least one further promoter and an output sequence, wherein each of said promoters comprises a transcription start site and/or is suitable for initiating transcription, wherein the initiation of transcription from the first promoter is enabled by a first transcriptional regulatory state and the initiation of transcription from each of said further promoter(s) is enabled by a respective further transcriptional regulatory state, and wherein said DNA construct yields an effective amount of an output RNA in a eukaryotic cell, when said first transcriptional regulatory state and/or any of the respective further transcriptional regulatory states is present in said cell, wherein said output RNA comprises a sequence corresponding to said output sequence. Furthermore, the invention relates to medical and/or diagnostic uses of the inventive DNA construct of the invention, e.g., for detecting, killing and/or manipulating different types of eukaryotic target cells in a subject and/or in a tissue sample.