Microbial inactivation of Pseudomonas putida and Pichia pastoris using gene silencing.
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Antisense deoxyoligonucleotide (ASO) gene silencing was investigated as a potential disinfection tool for industrial and drinking water treatment application. ASOs bind with their reverse complementary mRNA transcripts thereby blocking protein translation. While ASO silencing has mainly been studied in medicine, it may be useful for modulating gene expression and inactivating microorganisms in environmental applications. In this proof of concept work, gene targets were sh ble (zeocin resistance) and todE (catechol-2,3-dioxygenase) in Pichia pastoris and npt (kanamycin resistance) in Pseudomonas putida. A maximum 0.5-fold decrease in P. pastoris cell numbers was obtained following a 120 min incubation with single-stranded DNA (ssDNA) concentrations ranging from 0.2 to 200 nM as compared to the no ssDNA control. In P. putida, a maximum 5.2-fold decrease was obtained after 90 min with 400 nM ssDNA. While the silencing efficiencies varied for the 25 targets tested, these results suggest that protein activity as well as microbial growth can be altered using ASO gene silencing-based tools. If successful, this technology has the potential to eliminate some of the environmental and health issues associated with the use of strong chemical biocides. However, prior to its dissemination, more research is needed to increase silencing efficiency and develop effective delivery methods.
SubjectColony Count, Microbial
Drug Resistance, Bacterial
Microbial Sensitivity Tests
Published Version (Please cite this version)10.1021/es901404a
Publication InfoGunsch, Claudia K; Morey, SJ; & Morse, TO (2010). Microbial inactivation of Pseudomonas putida and Pichia pastoris using gene silencing. Environ Sci Technol, 44(9). pp. 3293-3297. 10.1021/es901404a. Retrieved from http://hdl.handle.net/10161/4030.
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Theodore Kennedy Associate Professor
Dr. Gunsch’s research focuses characterizing and engineering environmental microbiomes. Students and postdoctoral associates in her group apply fundamental concepts from the fields of microbiology, genomics and bioinformatics to environmental engineering applications. Current research projects focus on exploring ballast water microbiomes, exploring correlations between microbial adaptation and evolution stemming from their exposure to contaminants, characterizing the fate of genetica