2020 Research in the Capitol

Title

Inactivation of pathogenicity genes of the corn pathogen Fusarium verticilloides using CRISPR gene editing

Presentation Type

Poster Presentation

Abstract

In molecular biology, CRISPR-Cas9 is a useful tool in gene editing. In this study, a target-specific CRISPR-Cas9 vector containing an RNA-guided Cas9 nuclease specific for the FUM1 gene will be constructed and inserted into Fusarium verticillioides protoplasts. Spores will be collected and used for protoplast production, and successful transformation of these protoplasts with the plasmid DNA will be detected through the use of hygromycin as a selective marker. The anticipated results for this study is that the FUM1 gene will be silenced in the transformed protoplasts. Strains of Fusarium verticillioides with a disrupted FUM1 gene fail to produce detectable levels of fumonisins, a known carcinogen. The transformed colonies will be detected through the use of hygromycin resistance, so the colonies that successfully grow on hygromycin plates will contain a mutated FUM1 gene. DNA sequencing will be used to confirm that a deletion in the FUM1 gene has occurred.

Start Date

24-3-2020 11:00 AM

End Date

24-3-2020 2:30 PM

Event Host

University Honors Programs, Iowa Regent Universities

Faculty Advisor

James Jurgenson

Department

Department of Biology

File Format

application/pdf

Electronic copy is not available through UNI ScholarWorks.

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Mar 24th, 11:00 AM Mar 24th, 2:30 PM

Inactivation of pathogenicity genes of the corn pathogen Fusarium verticilloides using CRISPR gene editing

In molecular biology, CRISPR-Cas9 is a useful tool in gene editing. In this study, a target-specific CRISPR-Cas9 vector containing an RNA-guided Cas9 nuclease specific for the FUM1 gene will be constructed and inserted into Fusarium verticillioides protoplasts. Spores will be collected and used for protoplast production, and successful transformation of these protoplasts with the plasmid DNA will be detected through the use of hygromycin as a selective marker. The anticipated results for this study is that the FUM1 gene will be silenced in the transformed protoplasts. Strains of Fusarium verticillioides with a disrupted FUM1 gene fail to produce detectable levels of fumonisins, a known carcinogen. The transformed colonies will be detected through the use of hygromycin resistance, so the colonies that successfully grow on hygromycin plates will contain a mutated FUM1 gene. DNA sequencing will be used to confirm that a deletion in the FUM1 gene has occurred.