Microbial inactivation of Pseudomonas putida and Pichia pastoris using gene silencing.
Abstract
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.
Type
Journal articleSubject
Colony Count, MicrobialDNA, Single-Stranded
Disinfection
Drug Resistance, Bacterial
Environmental Microbiology
Gene Silencing
Genetic Techniques
Kanamycin Resistance
Microbial Sensitivity Tests
Oligonucleotides, Antisense
Pichia
Pseudomonas putida
RNA, Messenger
Time Factors
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https://hdl.handle.net/10161/4030Published Version (Please cite this version)
10.1021/es901404aPublication Info
Morse, Thomas O; Morey, Sara J; & Gunsch, Claudia K (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.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
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 Precis

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