Effect of CRISPR/Cas9 Targets Associated with Iron Metabolism and Its Variation on Transcriptional Regulation of SHK-1 Cell Line as a Model for Iron Metabolism

• Published in: Fishes 2024-06, Vol.9 (6), p.198
• Main Authors: Dettleff, Phillip, Jin, Yehwa, Peñaloza, Carolina, Pulgar, Rodrigo, Sáez, Alejandro, Robledo, Diego, Escobar-Aguirre, Sebastian
• Format: Article
• Language: English
• Subjects: Aquaculture; cell lines; Cells; CRISPR; CRISPR-Cas systems; CRISPR-Cas9; Cytoplasm; Design; DNA; Efficiency; Gene editing; Gene expression; Gene regulation; Genes; genome editing; Genomes; gRNA; Iron; iron absorption; iron metabolism; Metabolism; Nucleic acids; Ribonucleic acid; RNA; RNA sequencing; RNA-seq; Salmon; sequence analysis; transcription (genetics); transcriptome; Transcriptomes; Transcriptomics; transferrin; Transferrins
• ISSN: 2410-3888; 2410-3888
• DOI: 10.3390/fishes9060198

In this study, we investigated the function of a gene associated with iron metabolism using CRISPR-Cas9 and RNA sequencing in SHK-1 salmon cells. Our objective was to understand how different guide RNA (gRNA) sequences against the transferrin gene tf could influence gene expression and cellular processes related to iron uptake. RNA-Seq analysis was performed to evaluate the transcriptomic effects of two distinct gRNA targets with high knock-out (KO) efficiencies for the targeted tf gene in the SHK-1 genome. Our results showed no significant differential expression in transferrin-related transcripts between wild-type and CRISPR-edited cells; however, there were major differences between their transcriptomes, indicating complex transcriptional regulation changes. Enrichment analysis highlighted specific processes and molecular functions, including those related to the nucleus, cytoplasm, and protein binding. Notably, different sgRNAs targeting tf might result in different mutations at DNA levels in SHK-1 salmon cells.