Investigating Gene Delivery Efficiency of Poly(β-amino ester) Derived From Poly(ethylene glycol) Diacrylate

Abstract

Poly(β-amino ester) (PβAE)-based polymers hold promise for bacterial gene transfer due to their ability to form stable complexes with genetic material and facilitate efficient delivery into bacterial cells. These polymers are easily modified to improve uptake and protect DNA or RNA from degradation, providing a safer, more controlled alternative to traditional methods like chemical transformation or electroporation. In this study, we evaluated the gene transformation efficiency of cationic PβAE polymer, which was synthesized through an aza-Michael addition reaction between piperazine and poly(ethylene glycol) diacrylate. Competent cells from E. coli strain DH5α were prepared with the optimized method using Ca²⁺ and Mg²⁺ ions. Different concentrations of the polymer were mixed with pRSET-EmGFP before transforming into competent cells through the heat shock method. Transformed cells were checked on medium containing ampicillin, and by using colony PCR, transformation efficiency was calculated. Based on our findings, we observed a successful transformation of the EmGFP gene in case of having the polymer. Furthermore, the PβAE at high concentrations (20–100 ng µL⁻¹) increased transformation efficiency more than twice as compared to the case of no polymer added. In conclusion, PβAE can effectively increase transformation efficiency.