Abstract BACKGROUND This study presents the development and application of magnetite/silver‐pDMAEMA‐PEG‐BUFII nanobioconjugates as an advanced plasmid delivery system in bacteria. These nanobioconjugates demonstrated superior efficiency compared to traditional methods in four challenging applications: (i) genomic editing in Gram‐positive bacteria; (ii) CRISPR/Cas9‐mediated antibiotic resistance control; (iii) large plasmid delivery; and (iv) transformation in complex environmental matrices such as soil. RESULTS Functionalization with pDMAEMA improved plasmid loading, while PEGylation enhanced stability, dispersity, and cellular uptake. The system enabled high‐efficiency transformation in Streptomyces JH010 , overcoming low conjugation efficiency observed with conventional approaches. In Escherichia coli , the nanobioconjugates facilitated effective CRISPR /Cas9‐based resensitization to antibiotics, achieving higher transformation and genome‐editing efficiencies than traditional competent‐cell methods. Furthermore, the nanobioconjugates achieved large plasmid delivery without requiring conjugation or phage‐based methods, thereby simplifying transformation procedures. In soil environments, plasmid transfer was significantly enhanced compared to transformation in chemically/competent cells, demonstrating potential for bioremediation and environmental applications. Characterization confirmed successful multi‐functionalization, enhanced colloidal stability, and low toxicity across bacterial models. The magnetic properties of the nanoparticles further enable potential recovery and reuse, reducing environmental impact. CONCLUSION These findings highlight the versatility and robustness of nanobioconjugate‐mediated plasmid delivery, providing an effective alternative for bacterial genetic engineering in medical, industrial, and environmental settings. Future research should focus on optimizing delivery conditions for diverse bacterial species, scaling up for industrial applications, and assessing long‐term impacts in complex biological systems. © 2025 The Author(s). Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI).
Páez‐Angarita et al. (Thu,) studied this question.