The Single-Stranded DNA Platform: A Potential Broad-Spectrum Vector for Editing Genes in Prokaryotes: ssDNA May Break Through the Difficulties of Genetic Manipulation on Non-Model Bacteria.

Single-stranded DNA (ssDNA) is an emerging nucleic acid tool with the potential to become a more efficient genetic delivery vector than traditional plasmids in prokaryotes. While conventional double-stranded DNA (dsDNA) vectors are easily recognized and attacked by bacterial defense systems, ssDNA's structural differences make it resistant to corresponding enzymatic recognition. Moreover, the presence of unpaired bases in ssDNA enables efficient binding with other molecules, allowing for regulated gene expression and targeted delivery of other nucleic acids or molecules. In this study, we investigated the natural forms of ssDNA and summarized its synthesis and introduction methods. By analyzing existing applications of ssDNA in eukaryotes, we demonstrated the strategies and advantages of applying it to genetic manipulation in prokaryotes.