This study evaluated the potential of an anti-lpxB pPNA to treat Acinetobacter baumannii infections alone or in combination with colistin. The objectives were to evaluate the lpxB gene in A. baumannii as a therapeutic target and propose anti-lpxB pPNAs to combat bacterial infection. The results showed that anti-lpxB pPNA reduced lpxB gene expression and that it had a better antimicrobial effect and synergy when combined with colistin. The conclusions determined that the lpxB gene is important for A. baumannii survival and that anti-lpxB pPNA inhibits its expression, demonstrating potential as a therapeutic approach.