Jennifer
Lee
Sponsor: Elizabeth Maga, Ph.D. Animal Science Gene editing in livestock is a valuable tool to advance desirable traits that benefit the agricultural field such as allergen-free milk production. Milk allergies affect 2-3% of children mainly due to the protein, β-lactoglobulin (BLG). One way to remove this allergen in milk is to render the gene non-functional using gene editing. Although CRISPR-Cas9 is a widely used gene editing tool due to its cost-effectiveness and efficiency, there are limited reports of its use in bovine embryonic stem cells (bESCs) as these cells were not readily available until recently. This project aims to optimize electroporation parameters in bESCs and use them to knock out the BLG gene with guide RNAs targeting exons 2 and 3. Electroporation, one method to deliver CRISPR-Cas9 reagents into bESCs, will first be optimized by comparing the transfection efficiency of different poring pulse voltages. Then, the editing efficiency of two guide RNAs will be analyzed using Tracking of Indels by Decomposition (TIDE). Successful completion of this research will provide proof of concept that the BLG gene can be knocked out in bESCs. Targeting BLG can also be used to enhance mastitis resistance by introducing genes for antimicrobial proteins into the locus in the future. Characterizing Thermal Stability of Genetically Diverse Rubiscos
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Jennifer Lee
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Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a carboxylase enzyme most well known for its role in the Calvin Benson Bassham (CBB) cycle, which fixes CO2 into biomass. Despite rubisco's critical role, its relatively slow catalytic rate has made it a prime candidate for targeted engineering efforts. However, only 38 clusters of rubiscos have been characterized, limiting the ability to properly evaluate the sequence space, fitness landscape, and biophysical limitations of the enzyme for engineering applications. Moreover, rubisco exhibits extensive phylogenetic diversity, encompassing multiple forms, such as Form I, prevalent in plants and cyanobacteria; Form II, found largely in bacteria; and Form III, present in archaea and some bacteria, that vary in subunit composition, metabolic contexts, and association with or absence from the canonical CBB cycle. To address these knowledge gaps, ongoing efforts identify and characterize novel rubiscos across the tree of life. In this study, ten genetically diverse rubiscos were successfully purified, revealing six new clades, and their thermal stability was characterized using a thermal shift assay. Notably, the newly characterized rubiscos generally had higher melting temperatures than the source organism's optimal growth temperature, indicating that rubiscos can maintain structural stability even above the conditions in which the host thrives. UC Davis 36 th Annual Undergraduate Research, Scholarship and Creative Activities Conference 163 The Impact of Alzheimer's Disease on the Onset and Progression of Type II Diabetes Julia Lee
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UC Davis / Chemistry / 2025
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Jennifer Lee