Finding efficient therapeutic strategies to fight antibiotic-resistant bacteria is a complicated affair specifically in the therapy of chronic bacterial infections related to hospital-acquired infections. Recently, three major antibacterial systems based on antisense RNA, CRISPR-Cas9, and metal/metal oxide nanoparticles particularly silver (Ag) nanoparticles have shown more effective antibacterial activity compared to conventional antibiotics. ROS generation, attachment to the cell membrane, disruption of bacterial envelop, inactivation of electron transport chain, decreasing the local pH, modulation of cell signaling, and denaturation of biological macromolecules such as proteins and nucleic acids have been found as the main antibacterial functions of Ag nanoparticles. Antisense RNA, a single-stranded RNA, can hybridize with complementary genes in messenger RNA (mRNA) followed by blockage translation of these genes into proteins. Moreover, CRISPR (clustered regularly interspaced short palindromic repeats) is a family of viral DNA sequences derived from bacteriophages, which can target and destroy foreign DNA by nuclease activity. There are 2 classes and 6 subtypes (I-VI) of CRISPR-Cas systems, which may be engineered as potential antibacterial agents to target specific sequences. Therefore, here, recent advances and challenges for the antibacterial application of these three therapeutic agents are presented (Source: journal of cellular, molecular, and biomedical reports).