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Peptide Nucleic Acids (PNAs) are single-stranded synthetic nucleic acids with a pseudopeptide backbone in lieu of the phosphodiester linked sugar and phosphate found in traditional oligos. PNA designed complementary to the bacterial Shine-Dalgarno or start codon regions of mRNA disrupts translation resulting in the transient reduction in protein expression. This study examines the use of PNA technology to interrupt protein expression in obligate intracellular  Rickettsia  sp. Their historically intractable genetic system limits characterization of protein function. We designed PNA targeting mRNA for  rOmpB  from  Rickettsia typhi  and  rickA  from  Rickettsia montanensis , ubiquitous factors important for infection. Using an  in vitro  translation system and competitive binding assays, we determined that our PNAs bind target regions. Electroporation of  R .  typhi  and  R .  montanensis  with PNA specific to  rOmpB  and  rickA , respectively, reduced the bacteria’s ability to infect host cells. These studies open the possibility of using PNA to suppress protein synthesis in obligate intracellular bacteria.

Disrupting Protein Expression with Peptide Nucleic Acids Reduces Infection by Obligate Intracellular Rickettsia