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Essential Gene(s) Targeted by Peptide Nucleic Acids Kills <i>Mycobacterium smegmatis</i>in culture and in Infected Macrophages

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Advances in Infectious Diseases 2021年第2期11卷 156-164页

作者： Md. Ariful Islam Mst. Minara Khatun Nammalwar Sriranganathan Stephen M. Boyle Center for One health Research Virginia-Maryland College of Veterinary Medicine Virginia Tech Blacksburg VA USA Department of Microbiology & hygiene Bangladesh Agricultural University Mymensingh Bangladesh

Background: Antisense peptide nucleic acids (PNAs) exhibit growth inhibitory effects on bacteria by inhibiting the expression of essential genes and could be promising therapeutic agents for treating bacterial infections. A study was carried out to determine the efficacy of several antisense PNAs in inhibiting extracellular and intracellular growth of Mycobacterium smegmatis. Methods: Six PNAs obtained from a commercial supplier were tested to evaluate the inhibitory effect on bacterial growth by inhibiting the expression of the following essential genes: inhA (a fatty acid elongase), rpsL (ribosomal S12 protein), gyrA (DNA gyrase), pncA (pyrazinamidase), polA (DNA polymerase I) and rpoC (RNA polymerase β subunit) of M. smegmatis. Each PNA was tested at 20 μM, 10 μM, 5 μM and 2.5 μM concentrations to determine whether they caused a dose dependent killing of M. smegmatis cultured in middlebrook 7h9 broth or in a J774A.1 murine macrophage cell line. Results: In middlebrook broth, the strong growth inhibitory effect against M. smegmatis was observed by PNAs targeting the inhA and rpsL genes at all four concentrations. The PNAs targeting the pncA, polA and rpoC genes were found to exhibit strong growth inhibition against M. smegmatis but only at 20 μM concentration. No growth inhibition of M. smegmatis was seen in pure culture when treated with PNAs targeting gyrA and a mismatch PNA targeting dnaG (DNA primase). All six PNAs showed killing of M. smegmatis in J774A.1 macrophage cell line that were statistically significant (p Conclusion: It may be concluded from this study that PNAs could be potential therapeutics for mycobacterial infections.

关键词： middlebrook 7h9 broth culture J774A.1 Murine Macrophage Cell Line Antisense Therapy Peptide Nucleic Acid Cell Penetrating Peptide Mycobacterium

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