Human Artificial Chromosome Vector Production in Xeno-Free Cells
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods fail to produce human artificial chromosome vectors in xeno-free environments with ensured safety for clinical applications, and they are not suitable for regenerative medicine due to contamination from nonhuman animal cells, especially when using mouse and chicken cells in vector construction.
Innovation Solution
A method is developed to produce human artificial chromosome vectors using human cells without nonhuman animal cells, involving the elimination of endogenous genes from human chromosomes to create a vector with a human centromere and telomere, allowing for the insertion of exogenous genes, thereby avoiding contamination and ensuring safety for clinical use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional methods using mouse A9 cells and chicken DT40 cells are used to produce human artificial chromosome vectors, then vector construction can be achieved, but contamination from nonhuman animal cells occurs and safety for clinical use is compromised
Solution Approach 1:
The invention extracts and removes the harmful nonhuman animal cell components (mouse A9 cells and chicken DT40 cells) from the vector construction process. By eliminating these foreign cell types and replacing them with human cells, the method removes the source of contamination while preserving the essential vector construction functionality.
Solution Approach 2:
The invention changes the biological parameter of cell origin from nonhuman (mouse and chicken) to human. This parameter change fundamentally alters the safety profile of the vector production method, making it suitable for clinical applications by ensuring xeno-free conditions.
2Reliability
If endogenous genes are retained in human chromosomes used for vector construction, then chromosome functionality is maintained, but the vector cannot be used for regenerative medicine due to potential harmful effects
Solution Approach 1:
The invention extracts and removes endogenous genes from the human chromosome structure used for vector construction. By eliminating these native genetic elements while retaining the essential chromosome components (centromere, telomere, origin of replication), the method creates a safe vector platform for regenerative medicine applications.
Solution Approach 2:
The invention applies local quality modification by selectively removing endogenous genes from specific regions of the human chromosome while preserving other critical functional elements. This localized modification creates a chromosome vector that is both functional and safe for clinical use.
3Adaptability or versatility
If human cells are used exclusively for vector construction, then xeno-free environment is achieved, but the complexity of producing functional artificial chromosomes increases
Solution Approach 1:
The invention makes human cells universal for the entire vector construction process, replacing the need for multiple different cell types (mouse and chicken cells). This multi-functional use of human cells simplifies the overall process while achieving xeno-free conditions, as human cells can perform all necessary functions including chromosome maintenance and vector production.
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 the production of human artificial chromosome vectors in a completely xeno-free and safe manner, suitable for regenerative medicine and gene therapy, by eliminating endogenous genes from human chromosomes to create a vector with a human centromere and telomere, allowing for the insertion of exogenous genes, thereby avoiding contamination and ensuring safety for clinical use.
Implementation Method 1
Patent Literature 2 and 3 describe methods for site-directed DNA cleavage and homogenous integration in mammalian cells, such as human cells, using genome editing with CRISPR/Cas9.
Data Source
AI summary
Provided is a method for producing a human artificial chromosome vector with high safety by using a human cell. The method for producing a human cell comprising a human artificial chromosome vector comprises: substantially eliminating endogenous genes of long-arm and short-arm in a disomic human cell containing a pair of the homologous human chromosomes or a trisomic human cell containing trisomy of the homologous human chromosomes, from one of two chromosomes of the disomy or one or two of three chromosomes of the trisomy, thereby producing a cell population containing a human cell containing a human artificial chromosome vector comprising long-arm moiety and short-arm moiety substantially containing no endogenous gene, a human centromere, and a telomere; and collecting the human cell containing the human artificial chromosome vector from the cell population. As well as, the human cell comprises the human artificial chromosome vector.