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.





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Morse, Thomas O, Sara J Morey and Claudia K Gunsch (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 https://hdl.handle.net/10161/4030.

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Claudia K. Gunsch

Professor in the Department of Civil and Environmental Engineering

Claudia Gunsch is a Professor of Civil and Environmental Engineering and holds secondary appointments in the Nicholas School of the Environment and the Department of Biomedical Engineering. She joined the Duke Faculty in 2004 after obtaining her PhD from the University of Texas at Austin, her MS from Clemson University and her BS from Purdue University. Currently, she serves as the Director for PreMiEr, the National Science Foundation Engineering Research Center for Precision Microbiome Engineering which is a joint venture between Duke University, North Carolina A&T State University, North Carolina State University, the University of North Carolina – Chapel Hill and the University of North Carolina – Charlotte. She also serves as an Associate Director for the Duke Microbiome Center. Previous leadership roles include serving as Associate Dean for Research and Infrastructure for the Pratt School of Engineering (2021-2022), Associate Vice Provost for Faculty Advancement (2019-2021) and as the Director of IBIEM (Integrative Bioinformatics for Investigating and Engineering Microbiomes), a joint graduate training program between Duke and North Carolina A&T State University (2015-2021).

Dr. Gunsch’s research bridges environmental engineering and molecular biotechnology. Current research foci include investigating the ecological impacts of emerging contaminants on environmental microbiomes, developing microbiome engineering approaches for bioremediation, studying microbial evolution following exposure to anthropogenic contaminants and developing innovative water treatment technologies. Her work has been funded in excess of $36 million by the National Science Foundation, US Environmental Protection Agency, National Institute for Environmental Health and Safety as well as state funding agencies and private industry.  Since becoming a faculty member, she has served as the primary mentor for 28 graduate students (8 MS and 20 PhD), 34 undergraduate students and 8 postdoctoral associates. She has been recognized for her research, teaching and service activities with several awards including the 2009 National Science Foundation Faculty Early Career Development Award, 2013 Langford Lectureship Award, 2016 Capers and Marion McDonald Award for Excellence in Mentoring and Advising and the 2016 American Society of Civil Engineers (ASCE) Walter L. Huber Civil Engineering Research Prize. Dr. Gunsch was also named ASCE Environmental & Water Resources Institute Fellow in 2022, Bass Fellow in 2016 and Fellow of the National Academy of Engineering for the United States Frontiers of Engineering in 2011 as well as the Indo-American Frontiers of Engineering in 2014.

She currently serves as Editor in Chief for Biodegradation. She is also a member of the Editorial Board for npj Clean Water and Industrial Biotechnology.  She serves on the Association of Environmental Engineering and Science Professors (AEESP) Board of Directors and has previously held several leadership roles within the Environmental & Water Resources Institute (EWRI).  Most recently, she served as the Environmental Council representative to the Technical Executive Committee to EWRI. 

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