A wholly new application
“IMPRINT represents an entirely new use of TXTL. While TXTL is widely employed for various purposes, including producing hard-to-express proteins or as affordable diagnostic tools, it has not previously been utilized to overcome barriers to DNA transformation in bacteria,” says Chase Beisel, head of the RNA Synthetic Biology department at the HIRI and professor at the JMU Medical Faculty. He spearheaded the study in collaboration with researchers from North Carolina State University (NC State) in Raleigh, USA. Their findings were published today in the journal Molecular Cell.
Compared to existing methods, IMPRINT offers speed and simplicity: “Current approaches require either laboriously purifying individual DNA methyltransferases or expressing them in E. coli, which often proves cytotoxic,” says Justin M. Vento, first author of the study who completed the work as a PhD student in the Department of Chemical and Biomolecular Engineering at NC State. “These methods can take days to weeks and only reconstitute a fraction of the bacterium's methylation pattern.”
The researchers demonstrated that IMPRINT could express a diverse array of DNA methyltransferases. Furthermore, these enzymes could be combined to recreate complex methylation patterns. This greatly enhanced DNA transformation in bacteria such as the pathogen Salmonella and the probiotic Bifidobacteria, including a challenging-to-transform strain of the latter, less-studied bacterium.
The basis for new antibiotics and cell-based therapies
The potential applications in modern medicine and research are extensive: IMPRINT can improve DNA transformation in clinical isolates of bacterial pathogens and in bacteria that combat infections, such as commensal bacteria or those producing antibacterial compounds. Genetic modification of these microbes could lead to new classes of antibiotics and cell-based therapies.
The research team aims to expand the use of IMPRINT: “We want to make a wide variety of bacterial pathogens genetically tractable for research,” Beisel says. He hopes that IMPRINT will be widely adopted by the research community: “Until now, certain bacteria have been favored as models simply because they are easier to genetically manipulate. We are hopeful that, by using IMPRINT, researchers will be able to focus on the most important bacterial strains, such as those with increased virulence or antibiotic resistance,” Beisel concludes.
Reference: Vento JM, Durmusoglu D, Li T, et al. A cell-free transcription-translation pipeline for recreating methylation patterns boosts DNA transformation in bacteria. Mol Cell. 2024:S1097276524004817. doi: 10.1016/j.molcel.2024.06.003
This article has been republished from the following materials. Note: material may have been edited for length and content. For further information, please contact the cited source. Our press release publishing policy can be accessed here.
