Date of Publication
2024
Document Type
Master's Thesis
Degree Name
Master of Science in Chemistry
Subject Categories
Chemistry
College
College of Science
Department/Unit
Chemistry
Thesis Advisor
Hilbert D. Magpantay
Abstract/Summary
Daphnia magna is a valuable model organism in environmental toxicology, but efficient gene-editing tools are crucial for elucidating gene function. This study evaluated a novel Cas9-GFP expressing D. magna strain for CRISPR-based gene editing. We assessed the transgenic phenotype by comparing growth rates and fecundity with wild-type strain, finding minimal impact on overall fitness. Cas9 expression was confirmed in embryos at 9 hours post-development. To optimize gene editing efficiency, we targeted the scarlet gene (DapmaST), responsible for eye pigmentation, using both capped and 2'-O-methylated gRNAs. While capped gRNA resulted in partial loss of eye pigmentation, methylated gRNA achieved complete loss, demonstrating superior editing efficacy. This study validates the functionality of our Cas9-GFP expressing D. magna model and highlights the importance of gRNA modification in enhancing CRISPR efficiency. Our findings establish an improved system for loss-of-function studies in D. magna, advancing its utility in environmental and genetic research.
Abstract Format
html
Language
English
Format
Electronic
Keywords
Daphnia magna; CRISPR-associated protein 9; fluorescence microscopy; Gene editing
Recommended Citation
Lumio, R. T. (2024). Phenotype and gene-editing evaluation of Cas9-GFP expressing daphnia magna. Retrieved from https://animorepository.dlsu.edu.ph/etdm_chem/21
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Embargo Period
9-5-2025
