Impact of GAUT1 Gene Knockout on Cell Aggregation in Arabidopsis thaliana Suspension Culture.

Abstract

The development of efficient producers of recombinant pharmaceuticals based on plant cell suspension cultures is a pressing challenge in modern applied science. A primary limitation of plant cell cultures is their relatively low yield of the target protein. One strategy to enhance culture productivity involves reducing cell aggregation. In order to minimize cell-to-cell adhesion in culture, we used Cas9 endonuclease to knock out the GAUT1 gene, which is a key gene of pectin biosynthesis in the genome of Arabidopsis thaliana. The resulting knockouts exhibited altered phenotypes and were unable to form viable plants. The suspension cell culture induced from seedlings bearing a homozygous deletion in the GAUT1 gene displayed darker coloration and an increased number of large aggregates compared to the control. The biomass accumulation rate showed no difference from the control, while the level of recombinant GFP protein accumulation was significantly reduced. Thus, our findings indicate that disruptions in pectin synthesis and the formation of larger aggregates in the suspension cell culture adversely affect the accumulation of the target recombinant protein. Alternative targets should be sought to reduce cell aggregation levels in plant cell cultures through genome editing.