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A Novel Method for Antisense Oligonucleotide Gene Expression Manipulation in Toxigenic Cyanobacterial Species, Microcystis Aeruginosa.

Record Type:	Language materials, manuscript : Monograph/item
Title/Author:	A Novel Method for Antisense Oligonucleotide Gene Expression Manipulation in Toxigenic Cyanobacterial Species, Microcystis Aeruginosa./
Author:	Velkme, Erik J.
Description:	1 online resource (75 pages)
Notes:	Source: Masters Abstracts International, Volume: 82-09.
Contained By:	Masters Abstracts International82-09.
Subject:	Microbiology. -
Online resource:	click for full text (PQDT)
ISBN:	9798582533528

A Novel Method for Antisense Oligonucleotide Gene Expression Manipulation in Toxigenic Cyanobacterial Species, Microcystis Aeruginosa.
Velkme, Erik J.

A Novel Method for Antisense Oligonucleotide Gene Expression Manipulation in Toxigenic Cyanobacterial Species, Microcystis Aeruginosa. - 1 online resource (75 pages)

Source: Masters Abstracts International, Volume: 82-09.

Thesis (M.S.)--Southern Illinois University at Carbondale, 2020.

Includes bibliographical references

Algal blooms caused by toxigenic cyanobacterial species are an increasing economic burden globally, as high anthropogenic inputs of nitrogen and phosphorous, coupled with rising levels of atmospheric CO2, promote eutrophication and enhance bloom proliferation. Of the freshwater bloom forming species, Microcystis aeruginosa has garnered the most attention due to the production of toxic secondary metabolites known as microcystins. These cyclic peptides are potent eukaryotic protein phosphatase 1 and 2A inhibitors, and can induce hepatic damage if concentration levels reach above the World Health Organization level of 1 µg/L. Current mitigation strategies of water column disruption or by use of broad acting chemicals, are limited in their range and may cause unwanted off target effects to the surrounding biota.Antisense oligonucleotides are short single-stranded DNA polymers that hybridize with transcribed mRNA, and suppress translation of protein products through steric hindrance of ribosomes, or by RNAse H degradation of the DNA/RNA bound complex. While antisense oligonucleotide applications have proven successful in the pharmaceutical industry, their potential remains largely unexplored in environmental contexts. For this reason, we investigated the knockdown of microcystin synthetase gene cluster mcyE in M. aeruginosa. We found that ionic charge neutralization coupled with heat shock were effective chemical competence based methods for delivery, mcyE transcript abundance in cells treated with phosphodiester linked antisense oligonucleotides significantly decreased in RT-qPCR analysis, and production of intracellular microcystin significantly decreased over a 24 hour period (-1.9 fg/cell). This work demonstrates a novel proof of concept for the potential use of exogenous antisense oligonucleotides to target M. aeruginosa in harmful algal bloom occurrences.

Electronic reproduction.
Ann Arbor, Mich. :
ProQuest,
2024

Mode of access: World Wide Web

ISBN: 9798582533528Subjects--Topical Terms:

591510
Microbiology.
Subjects--Index Terms:

Antisense oligonucleotidesIndex Terms--Genre/Form:

554714
Electronic books.

A Novel Method for Antisense Oligonucleotide Gene Expression Manipulation in Toxigenic Cyanobacterial Species, Microcystis Aeruginosa.
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520 $a Algal blooms caused by toxigenic cyanobacterial species are an increasing economic burden globally, as high anthropogenic inputs of nitrogen and phosphorous, coupled with rising levels of atmospheric CO2, promote eutrophication and enhance bloom proliferation. Of the freshwater bloom forming species, Microcystis aeruginosa has garnered the most attention due to the production of toxic secondary metabolites known as microcystins. These cyclic peptides are potent eukaryotic protein phosphatase 1 and 2A inhibitors, and can induce hepatic damage if concentration levels reach above the World Health Organization level of 1 µg/L. Current mitigation strategies of water column disruption or by use of broad acting chemicals, are limited in their range and may cause unwanted off target effects to the surrounding biota.Antisense oligonucleotides are short single-stranded DNA polymers that hybridize with transcribed mRNA, and suppress translation of protein products through steric hindrance of ribosomes, or by RNAse H degradation of the DNA/RNA bound complex. While antisense oligonucleotide applications have proven successful in the pharmaceutical industry, their potential remains largely unexplored in environmental contexts. For this reason, we investigated the knockdown of microcystin synthetase gene cluster mcyE in M. aeruginosa. We found that ionic charge neutralization coupled with heat shock were effective chemical competence based methods for delivery, mcyE transcript abundance in cells treated with phosphodiester linked antisense oligonucleotides significantly decreased in RT-qPCR analysis, and production of intracellular microcystin significantly decreased over a 24 hour period (-1.9 fg/cell). This work demonstrates a novel proof of concept for the potential use of exogenous antisense oligonucleotides to target M. aeruginosa in harmful algal bloom occurrences.
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