Fecha de publicación: -- Fuente: PubMed "plant biotechnology" Acta Biochim Biophys Sin (Shanghai). 2026 Nov 9. doi: 10.3724/abbs.2026193. Online ahead of print.ABSTRACTPlant-based expression systems have emerged as low-cost, scalable, and safe alternatives for producing therapeutic recombinant proteins. Among them, Nicotiana benthamiana stands out as a versatile chassis due to its rapid growth, high biomass, efficient transient expression, and weak RNA silencing. In this review, we systematically summarize the key technical procedures and recent advances in the expression of therapeutic recombinant proteins in N. benthamiana, covering gene design and subcellular targeting, expression vector engineering and delivery, host engineering for enhanced yield, and cultivation optimization. We discuss strategies to address major bottlenecks, including codon optimization, endoplasmic reticulum retention, protease knockout, glycoengineering to eliminate immunogenic plant-specific glycans, and the use of viral-based transient expression systems such as magnICON, pEAQ-HT, and geminiviral replicons. Additionally, we highlight emerging approaches such as CRISPR-based multiplexed chassis engineering, artificial intelligence-assisted vector design, and hydroponic cultivation for scalable production. Despite significant progress, challenges remain in achieving coordinated high-level expression of multiple proteins and reducing downstream processing costs. This review provides a comprehensive framework for understanding the current state and future directions of plant molecular farming in N. benthamiana, emphasizing its irreplaceable value for rapid emergency response, personalized vaccines, and safe biopharmaceutical manufacturing.PMID:43002214 | DOI:10.3724/abbs.2026193