An effective method to enhance transient gene expression by transfecting high-density CHO cells.

ABSTRACT: Chinese hamster ovary (CHO) cells are the predominant host for recombinant protein production. While transient gene expression (TGE) systems provide a faster, more cost-effective alternative to stable gene expression (SGE), their application is often hampered by low protein yields. Here, we optimized TGE systems based on high-density suspension transfection using 40 kDa polyethylenimine (PEI) by engineering vector regulatory elements (Kozak, polyA, introns, leader) and optimizing transfection conditions. Vector optimization led to a substantial enhancement in recombinant protein, with EGFP and SEAP levels increased by 5.33-fold and 4.13-fold, respectively (p < 0.05). By applying the optimized vector in combination with identified optimal conditions, a DNA concentration of 4 µg/mL, a DNA: P

SUBMITTER: Fu YS 

PROVIDER: S-EPMC12748440 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature