[Strategies for Optimizing Recombinant Protein Synthesis in Plant Cells: Classical Approaches and New Directions]

Area of Science:

• Biotechnology
• Molecular Biology
• Plant Science

Background:

• Plant expression systems offer a cost-effective alternative for producing pharmacologically significant proteins, combining bacterial simplicity with eukaryotic post-translational modifications.
• A key limitation of plant expression systems is the relatively low yield of target recombinant proteins compared to bacterial systems.
• Gene silencing, often caused by random insertion into heterochromatin, further hinders efficient recombinant protein production in plants.

Purpose of the Study:

• To review methods for enhancing recombinant gene expression in plant systems over the past two decades.
• To explore strategies for preventing gene silencing in plant expression systems.
• To introduce CRISPR/Cas technologies as a novel approach for increasing gene expression by targeted insertion into transcriptionally active regions.

Main Methods:

• Review of literature on methods to increase recombinant gene expression and prevent silencing in plant systems.
• Analysis of gene silencing mechanisms, including heterochromatin insertion.
• Examination of CRISPR/Cas technologies for directed gene insertion into specific plant genome loci.

Main Results:

• Various methods have been developed to improve recombinant protein yield and prevent silencing in plant expression systems.
• CRISPR/Cas technologies offer a new paradigm for enhancing gene expression through precise genomic integration.
• Identifying and utilizing transcriptionally active regions, such as housekeeping gene loci, is crucial for high-level expression.

Conclusions:

• Targeted insertion of recombinant genes into transcriptionally active plant genome regions, facilitated by CRISPR/Cas, significantly enhances protein yield.
• Understanding the organization of housekeeping genes provides promising targets for integrating pharmaceutical protein genes.
• Plant expression systems, when optimized with advanced techniques, represent a powerful platform for biopharmaceutical production.