Phenotype and gene-editing evaluation of Cas9-GFP expressing daphnia magna
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...
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| Format: | text |
| Language: | English |
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Animo Repository
2024
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| Online Access: | https://animorepository.dlsu.edu.ph/etdm_chem/21 https://animorepository.dlsu.edu.ph/context/etdm_chem/article/1021/viewcontent/2024_Lumio_Phenotype_and_Gene_editing_Evaluation_of_Cas9_GFP_Expressing_Daph.pdf |
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| Institution: | De La Salle University |
| Language: | English |
| 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. |
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