Hospital-associated infections – predominantly those caused by Enterococcus faecalis, a bacteria found in the human gut – can lead to a variety of multidrug-resistant, life-threatening complications. Thankfully, researchers at the Singapore-MIT Alliance for Research and Technology (SMART) have developed a new tool that can help understand and prevent physiological behaviours of bacteria such as E. faecalis, including biofilm development and drug resistance.

The SMART researchers, specifically from the Antimicrobial Resistance (AMR) Interdisciplinary Research Group, designed an easily modifiable genetic technique using famed CRISPRi technology that allows rapid and efficient silencing of E. faecalis genes to prevent infection and antibiotic resistance.

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The new tool is believed to be valuable in investigation of a wide range of aspects of enterococcal biology and pathogenesis, host-bacterium interactions, and interspecies communication. The genetic technique can also be scaled up to simultaneously silence multiple bacterial genes or perform full-genome studies for novel antimicrobial combinatorial therapies.

“Infections caused by E. faecalis are usually antibiotic tolerant and more difficult to treat, rendering them a significant public health threat,” says Dr. Irina Afonina, Postdoctoral Associate at SMART AMR.

“The system we designed enables us to easily interrogate various stages during the biofilm developmental cycle of E. faecalis,” adds SMART AMR Principal Investigator Kimberly Kline. “Bacterial biofilms are clusters of bacteria that are enclosed in a protective, self-produced matrix – by selectively silencing certain genes in pre-formed, mature biofilms, we can erode the biofilm and force it to disperse.”

Identifying the genes that are involved in these bacterial processes could help with discovery of new drug targets or propose antimicrobial strategies to treat E. faecalis infections and possibly overcome antimicrobial resistance, finishes Dr. Afonina.

SMART’s inducible CRISPRi system was co-developed by Nanyang Technological University (NTU) and supported by the National Research Foundation (NRF) Singapore under its Campus for Research Excellence And Technological Enterprise (CREATE) programme.