Disrupting protein expression with Peptide Nucleic Acids reduces infection by obligate intracellular Rickettsia

PloS One
Rebecca S PelcShane M Ceraul

Abstract

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.

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Citations

Dec 17, 2015·Infection and Immunity·Anna J Hooppaw, Derek J Fisher
Jun 24, 2017·Frontiers in Cellular and Infection Microbiology·Pratibha SharmaYasuko Rikihisa
Mar 31, 2021·Pathogens and Disease·Jon McGinn, Rebecca L Lamason

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Methods Mentioned

BETA
dissection
scraping
dot blot
Infection

Software Mentioned

GeneTools
BLASTn
Excel
QCapture Pro

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