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DNA vaccines represent a promising approach given their stability, safety, design flexibility and cost\u2010effectiveness for rapid production. In this study, two plasmid backbones, namely the commercial pVAX1 and the newly designed PLASMIVAX (containing additional CpG motifs), were tested to develop two DNA vaccines targeting the MrkA fimbrial subunit from\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>K. pneumoniae<\/jats:italic>\n                    <\/jats:styled-content>\n                    . MrkA is highly conserved among\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>K. pneumoniae<\/jats:italic>\n                    <\/jats:styled-content>\n                    and was expressed and secreted in\u00a0vitro, fused to immunostimulatory elements, by both DNA vaccines following transfection of HEK\u2010293T cells. Immunogenicity was assessed in mice by measuring antibody levels (IgM, IgG, IgG1, IgG2a and IgG3) after intramuscular immunization with three doses of 50\u2009\u03bcg of each DNA vaccine. Efficacy of protection was established in a sepsis infection model. The pVAX1\u2010MrkA DNA vaccine elicited the most robust antibody response and protection against\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>K. pneumoniae<\/jats:italic>\n                    <\/jats:styled-content>\n                    infection (80%,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.0373), while the PLASMIVAX\u2010MrkA DNA vaccine protected at a lower rate (30%,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.0163), consistent with lower post\u2010immunization antibody titers elicited by this vaccine. This study reveals that DNA plasmid properties strongly influence immunogenicity and vaccine efficacy and highlights the potential role of MrkA as a vaccine antigen and DNA platforms as a valuable tool for developing vaccines against multidrug\u2010resistant bacteria.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.70434","type":"journal-article","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T06:54:32Z","timestamp":1787554472000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Pre\u2010Clinical Development of Two\n                    <scp>DNA<\/scp>\n                    Vaccines Encoding\n                    <scp>MrkA<\/scp>\n                    of\n                    <i>Klebsiella pneumoniae<\/i>\n                    : Effect of Plasmid Backbone on Immunogenicity and Protective Efficacy Against 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