M.S. - Molecular Biosciences and Bioengineering
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Item UPGRADING BLACK SOLDIER FLY LARVAE MEAL FOR AQUATIC FEEDS USING A SUSTAINABLE MICROBIAL PROCESS(2024) Ochoa, Daniel M.; Su, Wei Wen; Molecular Biosciences and BioengineeringItem Evaluating the Role of EXOC5 in Early Folliculogenesis(2024) Young, Shaina Fei; Ward, Steven; Molecular Biosciences and BioengineeringItem Modifying Hawaiian Drosophila laboratory diet with material from the wild: Phenotypic response to microbial and nutritional variation(2024) Konicek, Kelli R.; Yew, Joanne; Molecular Biosciences and BioengineeringItem Germline BAP1 mutation alter the tumor immune microenvironment and impact the progression of Malignant Mesothelioma (MM)(2024) Tyagi, Garima; Carbone, Michele Dr; Molecular Biosciences and BioengineeringItem Elucidating and Rewiring Regulation of Terpenoid Biosynthesis in Yarrowia lipolytica Utilizing Renewable Lipid Feedstocks(2024) Maruwan, Jessica; Su, Wei-Wen; Molecular Biosciences and BioengineeringItem ESTABLISHING EFFICIENT PROTOPLAST GENOME EDITING SYSTEMS USING CRISPR/CAS9 FOR MAIZE BREEDING AND IMPROVEMENT(2024) Higa, Lauren Akemi; Du, Zhi-Yan (Rock); Molecular Biosciences and BioengineeringItem EFFECTS OF HAWAIIAN-GROWN COFFEE ON SERUM LEPTIN LEVELS AMONG HEALTHY INDIVIDUALS(2021) Stillwell, Nyan Crandall; Nerurkar, Pratibha V.; Molecular Biosciences and BioengineeringItem Development Of A Mesophyll Protoplast-based System For Gene Editing Of Papaya(University of Hawaii at Manoa, 2022) Elias, Marc Joseph; Christopher, David A.; Molecular Biosciences and BioengineeringPapaya (Carica papaya L.) is an economically important food crop grown in tropical and subtropical regions. The global gross agricultural production value for papaya reached $6.2 billion in 2020. Papaya and its byproducts are used in commercial production, cosmetics, and pharmaceutical industries, offering many health benefits. While papaya is important economically, it faces threats from microbial pathogens. Genetic engineering and traditional breeding methods have been used to produce papaya with improved resistance to some pathogens. However, few innate disease-resistance genes are present in the papaya genome, and novel gene introgression from closely related species has reported F1 sterility with low viability and vigor. CRISPR-Cas9 gene editing technology has enabled gene editing in papaya using the Agrobacterium-mediated transformation method. When introduced to the market, papaya developed using this transgenic method will face increased regulatory constraints. A non-transgenic DNA-free alternative is available using CRISPR-Cas9 ribonucleoprotein complexes. A high-yield protoplast isolation method, producing highly viable, durable protoplasts, was developed during this study. Papaya protoplasts were transfected with fluorescent reporter constructs using our papaya-optimized PEG-mediated transfection method. Transfected cells were shown to express and shuttle proteins to expected locations and organelles viably. A system for DNA-free gene editing in papaya was developed using PEG-mediated transfection to deliver CRISPR-Cas9 ribonucleoprotein complexes into papaya protoplasts. The albino phenotypic marker gene phytoene desaturase and the susceptibility gene MLO homolog mildew resistance locus O-like protein 6 were targeted during this study.Item Chemical Approaches To Improve The Manufacturability Of Disulfide-rich Peptide Toxins – Engineering A Stable And Selective Peptide Blocker Of The KCa1.1 Channel(University of Hawaii at Manoa, 2022) Delgado, Erick Medermel; Bingham, Jon-Paul; Biological EngineeringScorpion venoms represent an immense source of bioactive peptides which are potent and selective effectors of sodium (Na+), potassium (K+), chloride (Cl-), and calcium (Ca2+) ion channels. Many of these compounds boast premium advantages which qualify them as attractive molecular tools and pharmaceutical leads: they penetrate tissue effectively while mitigating off-target toxicity, are capable of selectively binding specific ion channel subtypes, alter channel function in a variety of desired ways: inhibit, potentiate, change the kinetics, etc. Unfortunately, many scorpion venom peptides feature unstable functional groups, are prone to aggregation during synthesis or oxidation, and require specific disulfide connectivity, all of which can complicate manufacturing and ultimately reduce synthetic yield. Using solid-phase peptide synthesis, native chemical ligation (NCL), and orthogonal bioconjugation, an effective bioengineering approach was developed for a stable and selective peptide blocker of the KCa1.1, calcium-activated potassium channel. This novel 37 amino acid Iberiotoxin (IbTX) analog, azido-IbTX-W14F, incorporates a mid-sequence Val16Ala mutation to facilitate thioester mediated NCL, homologous substitutions of oxidation prone Met29 and Tyr14 residues, and an orthogonally conjugated azido-Lys19 residue capable of fluorescent bioconjugation with a phosphine fluorophore. Azido-IbTX-W14F retained native-like bioactivity according to electrophysiology studies, exhibited increased solution stability, and was demonstrated to be amenable to chemoselective fluorescent bioconjugation. The resulting fluorescent-IbTX analog has potential as a diagnostic tool or as a drug lead for disease states mediated by KCa1.1.Item Flavonoid Metabolon Formation In Citrus Sinensis(University of Hawaii at Manoa, 2022) Knittel, David; Owens, Daniel; Molecular Biosciences and BioengineeringThe flavonoid metabolon has been previously theorized and researched, but little work has been done on the mapping of later pathway and periphery enzymes. This research has demonstrated novel characterizations of interactions among glucosyltransferases (GTs) and the primary flavonoid pathway as well as created rapid assays for the characterization of protein-protein interactions of multiple classes of enzymes. Additionally, several assays have been created for the analysis of membrane bound P450 class enzymes, specifically Flavone Synthase II (FNSII). These procedures will allow for the rapid assaying of a wide variety of enzymes alongside partner groups of various experience levels. The putative flavonoid metabolon structure of Citrus sinensis differs from previously examined Arabidopsis thaliana and may provide insight into the stabilizing effects of periphery pathway enzymes. Genetic differences between blond and blood cultivars of sweet orange were also identified as potential binding sequences in GT enzymes.