DNA Element Enhancing Foreign Gene Expression via Chromatin Modification
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Solution Overview
Problem
Current methods for producing therapeutic proteins in mammalian cells face challenges such as low productivity, high costs, and the need for large-scale cell cultures, primarily due to the 'position effect' of foreign gene integration, which affects transcription and is difficult to control.
Innovation Solution
The use of specific DNA elements, such as locus control regions (LCRs), scaffold/matrix attachment regions (S/MARs), and other regulatory sequences to enhance chromatin structure changes around the foreign gene locus, improving transcription and translation efficiency when combined with promoters in foreign gene expression vectors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a foreign gene is randomly integrated into the host chromosomal genome, then the foreign gene can be expressed, but the transcription is greatly affected by the position effect and productivity remains low
Solution Approach 1:
The patent introduces a Locus Control Region (LCR) as an intermediary DNA element between the foreign gene and the chromosomal integration site. This LCR acts as a mediator that protects the foreign gene from position effects by providing a controlled chromatin environment, thereby stabilizing transcription while maintaining high expression levels
Solution Approach 2:
The patent changes the chromatin structure parameters around the foreign gene integration site by introducing specific DNA elements (LCR, S/MAR). These elements modify local chromatin accessibility and nucleosome positioning, transforming the transcriptional environment from unstable to stable high-expression state
2Reliability
If cultured mammalian cells are used as host cells, then post-translational modification can be performed, but the growth rate is low and productivity is low
Solution Approach 1:
The patent changes the genetic parameters of the mammalian cell system by introducing optimized expression vectors containing LCR elements. This modifies the transcriptional and translational parameters, increasing protein production rate while preserving the cell's inherent post-translational modification capabilities
3Quantity of substance
If a large amount of foreign protein is produced for clinical trials, then sufficient product is available, but the cost is high due to low productivity
Solution Approach 1:
The patent applies preliminary action by pre-engineering the expression system with LCR elements and optimized promoters before large-scale production. This preliminary optimization of the genetic architecture enables high productivity from the outset, reducing the scale and cost of subsequent production for clinical trials
Data Source
AI summary
A method for stably achieving high expression of a foreign gene in mammalian cells using a novel DNA element is disclosed. More specifically, the present application discloses a DNA element which enhances the activation of transcription by changing the chromatin structure around a gene locus into which a foreign gene expression unit has been introduced.