Nucleic acid analogues are used in protein synthesis, as their effects on gene expression are confirmed, and play a crucial and important role during transcriptional and posttranscriptional processes in the cell. Due to the many remaining challenges associated with genetic engineering, current research mainly focuses on new materials such as peptide nucleic acids [PNA] and locked nucleic acids [LNA]. PNAs and LNAs have been developed to mimic the chemical characteristics of DNA and RNA and to show improved properties. They represent synthetic DN/RNA analogues in which the phosphodiester backbone is replaced with unchanged aminoethylglycine units, being very resistant to enzymatic degradation by proteases and nucleases. This review summarizes the application of nucleic acid analogues in the inhibition of gene expression in bacterial genomes, as well as the usage in the development of high-performance affinity biosensors.