Genome Cloning via Yeast Intermediary Host
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for cloning and manipulating large nucleic acids, such as genomes, are limited by size restrictions and incompatibility issues between different cell types, leading to toxicity and instability when transferring nucleic acids between organisms of different species or genera, particularly from prokaryotic to eukaryotic cells and back.
Innovation Solution
The development of methods and systems for transferring, modifying, and transplanting nucleic acids across different species by using host cells with strong genetic systems like yeast, allowing for the manipulation of nucleic acids greater than 300 kb, including whole genomes, and introducing host vectors to facilitate homologous recombination and modification, enabling the creation of new phenotypes and genomes not previously existing in nature.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional cloning methods are used to transfer nucleic acids between different species or genera, then the transfer process is simple, but the nucleic acid size is limited and incompatibility issues arise leading to toxicity and instability
Solution Approach 1:
The patent uses an intermediary host system (yeast or other eukaryotic cells) that is genetically compatible with both the donor and recipient organisms. This intermediary allows large nucleic acids to be transferred and maintained stably through controlled homologous recombination, avoiding the toxicity and instability problems that occur with direct transfer between incompatible species.
Solution Approach 2:
The patent changes the genetic parameters of the host cell by introducing modified host vectors with altered recombination signals and selection markers. This allows the host to accommodate and manipulate nucleic acids of varying sizes and from different species, expanding the adaptability range while maintaining stability through controlled recombination parameters.
2Adaptability or versatility
If large nucleic acids greater than 300 kb are transferred into host cells using conventional methods, then the genetic manipulation capability is enhanced, but toxicity and instability occur due to incompatibility between different cell types
Solution Approach 1:
The patent employs an intermediary host system with modified genetic tools that can accommodate large nucleic acids from different species. The host vectors are designed to mediate the transfer and integration process, reducing toxicity and instability by controlling the recombination events between the transferred nucleic acid and the host genome.
Solution Approach 2:
The patent performs preliminary modification of host vectors before introducing large nucleic acids. The host vectors are pre-engineered with appropriate recombination signals, selection markers, and regulatory elements that prepare the host system to accept and stably maintain large foreign nucleic acids, preventing toxicity and instability issues.
3Manufacturing precision
If host vectors are introduced to facilitate homologous recombination and modification of large nucleic acids, then the precision of genetic manipulation is improved, but the device complexity increases
Solution Approach 1:
The patent segments the host vector system into modular components including recombination signals, selection markers, and regulatory elements. This segmentation allows precise genetic manipulation by enabling independent optimization and combination of vector components, while the modular nature actually reduces overall system complexity through standardized interfaces and functions.
Solution Approach 2:
The patent designs host vectors with universal features that can accommodate different large nucleic acids from various species. The vectors contain universal recombination signals and selection systems that work across different genetic backgrounds, improving precision while reducing complexity by eliminating the need for custom vector design for each manipulation task.
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
Enables the successful transfer and modification of large nucleic acids across species, overcoming size limitations and incompatibility issues, allowing for the production of new phenotypes and genomes that were previously unattainable, with the ability to modify and engineer organisms for specific traits and products like biofuels and therapeutic applications.
Implementation Method 1
introducing the donor genome and a host vector into a heterologous host cell, wherein the donor genome and the host vector are optionally joined prior to introduction into the host cell
Data Source
AI summary
Compositions and methods are disclosed herein for cloning a donor genome in a heterologous host cell. In one embodiment, the donor genome can be further modified within a host cell. Modified or unmodified genomes can be further isolated from the host cell and transferred to a recipient cell. Methods disclosed herein can be used to alter donor genomes from intractable donor cells in more tractable host cells.