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Peptide Nucleic Acids (PNAs), synthetic DNA analogues with peptide-like backbones, act as thermically, chemically, and enzymatically stable sequence-specific agents capable of silencing essential bacterial genes. Through antisense mechanisms, PNAs bind bacterial mRNA or rRNA, blocking translation or ribosome assembly and thereby inducing species-specific growth inhibition. Their programmable design enables precise targeting of multidrug-resistant pathogens while sparing commensal microbiota. Recent advances, including \u03b3-modified backbones, cationic substitutions, and delivery platforms such as cell-penetrating peptides (CPPs), dendron conjugates, and nanoparticles, have improved solubility, stability, and cellular uptake. Studies show promising in vitro and, albeit less frequently, in vivo efficacy against both Gram-positive and Gram-negative bacteria, often with synergistic activity when combined with conventional antibiotics. Although challenges remain in delivery and large-scale production, PNAs represent a promising class of antimicrobials to combat AMR through targeted gene inhibition.<\/jats:p>","DOI":"10.3390\/ijms27031565","type":"journal-article","created":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T11:13:01Z","timestamp":1770289981000},"page":"1565","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Peptide Nucleic Acids (PNAs) in Antimicrobial Therapy: A Next Generation Strategy"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-8506-6388","authenticated-orcid":false,"given":"Antonia","family":"D\u2019Aniello","sequence":"first","affiliation":[{"name":"Institute of Crystallography, National Research Council of Italy, URT Caserta, Via Vivaldi 43, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7163-3978","authenticated-orcid":false,"given":"Annalisa","family":"Masi","sequence":"additional","affiliation":[{"name":"Institute of Crystallography, National Research Council of Italy, Strada Provinciale 35d, 9, Montelibretti, 00010 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5542-6920","authenticated-orcid":false,"given":"Concetta","family":"Avitabile","sequence":"additional","affiliation":[{"name":"Institute of Crystallography, National Research Council of Italy, URT Caserta, Via Vivaldi 43, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"del Monaco","sequence":"additional","affiliation":[{"name":"Institute of Crystallography, National Research Council of Italy, URT Caserta, Via Vivaldi 43, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5086-2459","authenticated-orcid":false,"given":"Michele","family":"Saviano","sequence":"additional","affiliation":[{"name":"Institute of Crystallography, National Research Council of Italy, URT Caserta, Via Vivaldi 43, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5279-3928","authenticated-orcid":false,"given":"Maria","family":"Moccia","sequence":"additional","affiliation":[{"name":"Institute of Crystallography, National Research Council of Italy, Strada Provinciale 35d, 9, Montelibretti, 00010 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,5]]},"reference":[{"key":"ref_1","unstructured":"(2025, October 16). 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