Improving transient protein expression in agroinfiltrated Nicotiana benthamiana

• Published in: The New phytologist Vol. 243; no. 3; pp. 846 - 850
• Main Authors: Beritza, Konstantina, Watts, Emma C., Hoorn, Renier A. L.
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.08.2024; John Wiley and Sons Inc
• Subjects: Agrobacterium tumefaciens; agroinfiltration; Biodegradation; Degradation; Endoplasmic reticulum; ER stress; Forum; gene silencing; Immune response; immunity; Inhibitors; Mutants; Nicotiana benthamiana; Protease; Protease inhibitors; Proteinase inhibitors; Proteins; proteolysis; Recombinants; RNA-mediated interference; transient expression; Viewpoints; Nicotiana benthamiana; transient expression; Agrobacterium tumefaciens; ER stress; agroinfiltration; gene silencing; immunity; proteolysis
• ISSN: 0028-646X; 1469-8137; 1469-8137
• DOI: 10.1111/nph.19894

Summary Agroinfiltration of Nicotiana benthamiana is routinely used in plant science and molecular pharming to transiently express proteins of interest. Here, we discuss four phenomena that should be avoided to improve transient expression. Immune responses can be avoided by depleting immune receptors and employing pathogen‐derived effectors; transcript degradation by using silencing inhibitors or RNA interference machinery mutants; endoplasmic reticulum stress by co‐expressing chaperones; and protein degradation can be avoided with subcellular targeting, protease mutants and co‐expressing protease inhibitors. We summarise the reported increased yields for various recombinant proteins achieved with these approaches and highlight remaining challenges to further improve the efficiency of this versatile protein expression platform.