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Development Of A Long-Read Sequencing Protocol To Assess The Precision And Efficacy Of Gene Editing For Duchenne Muscular Dystrophy, Landon Andrew Burcham
Development Of A Long-Read Sequencing Protocol To Assess The Precision And Efficacy Of Gene Editing For Duchenne Muscular Dystrophy, Landon Andrew Burcham
Graduate Theses and Dissertations
This work establishes a method for assessing on-target precision due to CRISPR-Cas9 gene editing, especially within the context of exon skipping therapy for Duchenne Muscular Dystrophy. The proposed method utilizes an Oxford nanopore long-read sequencing approach to sequence amplified regions of DNA that have been edited using CRISPR-Cas9. NIH3T3 and C2C12 cell lines were treated with a dual-guide CRISPR-Cas9 system, that targets and deletes exon 23 from the DMD gene in mouse samples. Deletion PCR revealed deletion of exon 23 in both DNA and cDNA samples. Additionally, sequencing using Oxford Nanopore revealed targeted exon 23 deletion as the most prevalent …
Developing Pennycress ( Thlaspi Arvense ) As A Biodiesel Feedstock Crop And Plant Model System, Michaela Mcginn
Developing Pennycress ( Thlaspi Arvense ) As A Biodiesel Feedstock Crop And Plant Model System, Michaela Mcginn
Theses and Dissertations
Thlapsi arvense L. (pennycress) is currently being developed as a profitable oilseed-producing winter annual cover crop with extreme cold tolerance and a rapid life cycle that can be grown on fallow farmland throughout the U.S. Midwest Corn Belt, controlling soil erosion and nutrient runoff while serving as an additional source of income for the American farmer without displacing food crops. The research comprising this dissertation demonstrates that pennycress can serve as a user-friendly model system highly similar to Arabidopsis thaliana, and is well-suited for both laboratory and field experimentation, being readily employable in existing growth facilities. After 10 generations of …