Nicotiana hairy roots for recombinant protein expression, where to start? A systematic review

Abstract

Hairy roots are a plant-tissue culture raised by Rhizobium rhizogenes infection (formerly known as Agrobacterium rhizogenes). Nowadays, these roots have been gaining more space in biotechnology due to their bene ts for the recombinant expression of valuables proteins; it includes simpli ed downstream processing, protein rhizosecretion, and scalability in bioreactors. However, due to methodological inconsistency among reports, the tissue platform is still a disruptive technology. In the current paper, we propose the rst step to overcome this issue through a systematic review of studies that employ Nicotiana hairy roots for recombinant expression. We conducted a qualitative synthesis of 36 out of 387 publications initially selected. Following the PRISMA procedure, all papers were assessed for exclusion and inclusion criteria. Multiple points of root culture were explored, including transformation methods, root growth curve, external additives, and scale-up with bioreactors to determine which approaches performed best and what is still required to achieve a robust protocol. The information presented here may help researchers who want to work with hairy roots in their laboratories trace a successful path to high recombinant expression.

Discontinuous Translocation of a Luciferase Protein beyond Graft Junction in Tobacco

“…Hairy roots are root organs transformed with the Ri plasmid. The characteristics of hairy roots include rapid growth and a high capacity for secondary metabolite and recombinant protein production 29 , 30 ) . Next, we studied whether or not the movement of LUC protein from the rootstock to the scion was strengthened by the hairy root phenotype.…”

“…In contrast, the effect of scion condition on NEP translocation was nearly negligible, since mentor grafting has quite a limited effect. Hairy roots are well known to grow vigorously and produce secondary metabolites and NEPs 29 , 30 ) . However, our results also suggest that hairy roots can systemically export NEPs more efficiently than normal roots.…”

Evaluation of led light on transgenic Nicotiana benthamiana production of a recombinant antibody fragment scFvanti-BAP1.

“…However, the costs of plant-based production are similar to the costs in animal cells due to the recombinant low-yield and expensive downstream processing. In this regard, several avenues have been explored with the potential to cheaper this process, with strategies to overcome gene silencing, handling of protein glycosylation, tissue culture adoption, and bioreactor scaling (Aragão et al, 2023;Donini and Marusic, 2019;Schillberg et al, 2019). The control of growing conditions such as temperature, light, pH; that allows the puri cation of the product and that also does not have high costs (Schillberg and Finnern, 2021).…”