HIRI researchers have developed a new method called IMPRINT to improve DNA transformation and genetic mutation in bacteria. Current genetic engineering methods are limited to a few bacterial species.
However, IMPRINT uses cell-free systems to enhance DNA transformation across many strains. This breakthrough, published in Molecular Cell, could lead to new ways of diagnosing, treating, and preventing infections, protecting crops, and creating eco-friendly chemical production.
A significant challenge in DNA transformation is the bacteria’s restriction-modification systems, which destroy foreign DNA that lacks its unique methylation pattern. Adding this pattern to DNA is complex and strain-specific.
To address this, researchers at HIRI and JMU developed IMPRINT. This new method replicates these methylation patterns using a cell-free transcription-translation system. This system produces the necessary enzymes to methylate DNA before introducing it into the target bacteria, enhancing DNA transformation.
IMPRINT is a new use of TXTL, typically used for protein production and diagnostics,” says Chase Beisel, head of the RNA Synthetic Biology department at HIRI and professor at JMU. He led the study with researchers from NC State. Their findings, published in Molecular Cell, show that IMPRINT is faster and simpler than existing methods.
It avoids the laborious purification of DNA methyltransferases and cytotoxicity issues in E. coli. IMPRINT can express various DNA methyltransferases to recreate complex methylation patterns, enhancing DNA transformation in bacteria like Salmonella and Bifidobacteria.
IMPRINT has many potential applications in medicine and research. It can improve DNA transformation in harmful and beneficial bacteria that fight infections, possibly leading to new antibiotics and cell-based therapies.
The research team aims to expand IMPRINT’s use to make more bacterial pathogens genetically manageable for research. Beisel hopes it will help scientists focus on crucial bacterial strains, like those with increased virulence or antibiotic resistance.
Journal reference:
- Justin M. Vento, Deniz Durmusoglu, et al., A cell-free transcription-translation pipeline for recreating methylation patterns boosts DNA transformation in bacteria. Molecular Cell. DOI: 10.1016/j.molcel.2024.06.003.