Shuttle Vectors for Automated Nucleic Acid Editing
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Solution Overview
Problem
Current methods for editing nucleic acids and shuttle vectors in heterologous cells are not compatible with automation due to low efficiencies and challenges with cell selection, making it difficult to edit exogenous polynucleotides, especially larger artificial chromosomes, in a cell-compatible manner.
Innovation Solution
The development of shuttle vectors and automated multi-module cell processing instruments that enable nucleic-acid guided nuclease editing of target polynucleotides in heterologous editing cells, allowing for the transfer and editing of exogenous polynucleotides using a system that includes inserting target polynucleotides into shuttle vectors, growing and concentrating cells, introducing editing vectors, and allowing editing to occur in a controlled environment within an automated instrument.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If manual methods are used for editing nucleic acids in heterologous cells, then flexibility and adaptability are maintained, but automation compatibility is poor and efficiency is low
Solution Approach 1:
The cell editing process is divided into distinct functional modules: cell growth module, cell concentration module, transformation module, and editing module. Each module performs a specific function in the workflow, enabling automated execution while maintaining flexibility for different cell types and editing applications.
Solution Approach 2:
The multi-module cell processing instrument is designed to accommodate various cell types (bacterial, yeast, mammalian) and different editing scenarios through a universal platform. The system can handle diverse polynucleotide sizes and types, making it applicable to broad research, production, and therapeutic contexts.
2Quantity of substance
If larger artificial chromosomes are edited, then payload size capacity increases, but cell compatibility and maintenance become more difficult
Solution Approach 1:
Shuttle vectors serve as intermediary carriers that can accommodate large polynucleotide inserts and facilitate their transfer between different cell types. The shuttle vector system enables maintenance of large artificial chromosomes in heterologous cells by leveraging the vector's ability to replicate and stabilize large DNA payloads across different cellular environments.
3Productivity
If automated multi-module instruments are implemented, then productivity and consistency improve, but device complexity increases
Solution Approach 1:
The instrument is segmented into independent modules (growth module, concentration module, transformation module, editing module), each with dedicated functions. This modular architecture enables automated high-throughput processing while keeping individual module complexity manageable and allowing flexible configuration for different applications.
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
This approach enables efficient and automated nucleic-acid guided nuclease editing of exogenous polynucleotides across various cell types, including bacterial, yeast, and mammalian cells, facilitating downstream applications such as re-introduction of edited polynucleotides into source organisms, thereby enhancing research, production, and therapeutic applications.
Implementation Method 1
Certain nucleases create site-specific double-strand breaks at target regions in the genome, which can be repaired by homologous recombination, resulting in targeted edits.
Implementation Method 2
Certain nucleases create site-specific double-strand breaks at target regions in the genome, which can be repaired by homologous recombination, resulting in targeted edits.
Implementation Method 3
There are various and diverse methods for delivering exogenous polynucleotides from the genome of one organism to another
Implementation Method 4
concentrating and rendering electrocompetent the heterologous editing cells in the cell concentration module
Implementation Method 5
introducing the library of shuttle vectors into the heterologous editing cells in a transformation module
Data Source
AI summary
The present disclosure provides shuttle vectors for editing exogenous polynucleotides in heterologous live cells, as well as automated methods, modules, and multi-module cell editing instruments and systems for performing the editing methods.