ArticleScientific reports2021
An efficient vector-based CRISPR/Cas9 system in an Oreochromis mossambicus cell line using endogenous promoters.
Article in Scientific reports, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 25 citations in OpenAlex.
- Efficient Gene Editing in Fish Primary Germline Stem Cells.International journal of molecular sciences · 2026Article
- Efficient genome editing in a Mozambique tilapia cell line using CAS ribonucleoprotein complexes.Scientific reports · 2026Article
- Comparative Analysis of CRISPR/Cas9 Delivery Methods in Marine Teleost Cell Lines.International journal of molecular sciences · 2025Article
- Electroporation-based CRISPR/Cas9 Gene Editing in Haliotis Discus Hannai.Marine biotechnology (New York, N.Y.) · 2025Article
- Comparative Analysis of Promoter Activity in Crassostrea gigas Embryos: Implications for Bivalve Gene Editing.Marine biotechnology (New York, N.Y.) · 2024Article
- Exploring the Use of Alternative Promoters for Enhanced Transgene and sgRNA Expression in Atlantic Salmon Cells.Marine biotechnology (New York, N.Y.) · 2024Article
- CRISPR/Cas9 Technology for Enhancing Desirable Traits of Fish Species in Aquaculture.International journal of molecular sciences · 2024Review
- An Efficient Vector-Based CRISPR/Cas9 System in Zebrafish Cell Line.Marine biotechnology (New York, N.Y.) · 2024Article
- Review
- Genome editing in East African cichlids and tilapias: state-of-the-art and future directions.Open biology · 2023Review
- Removal of evolutionarily conserved functional MYC domains in a tilapia cell line using a vector-based CRISPR/Cas9 system.Scientific reports · 2023Article
- Molecular Characterization of U6 Promoters from Orange-Spotted Grouper (Epinephelus coioides) and Its Application in DNA Vector-Based RNAi Technology.Marine biotechnology (New York, N.Y.) · 2023Article
- Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR/Cas9 gene editing is effective in manipulating genetic loci in mammalian cell cultures and whole fish but efficient platforms applicable to fish cell lines are currently limited. Our initial attempts to employ this technology in fish cell lines using heterologous promoters or a ribonucleoprotein approach failed to indicate genomic alteration at targeted sites in a tilapia brain cell line (OmB). For potential use in a DNA vector approach, endogenous tilapia beta Actin (OmBAct), EF1 alpha (OmEF1a), and U6 (TU6) promoters were isolated. The strongest candidate promoter determined by EGFP reporter assay, OmEF1a, was used to drive constitutive Cas9 expression in a modified OmB cell line (Cas9-OmB1). Cas9-OmB1 cell transfection with vectors expressing gRNAs driven by the TU6 promoter achieved mutational efficiencies as high as 81% following hygromycin selection. Mutations were not detected using human and zebrafish U6 promoters demonstrating the phylogenetic proximity of U6 promoters as critical when used for gRNA expression. Sequence alteration to TU6 improved mutation rate and cloning efficiency. In conclusion, we report new tools for ectopic expression and a highly efficient, economical system for manipulation of genomic loci and evaluation of their causal relationship with adaptive cellular phenotypes by CRISPR/Cas9 gene editing in fish cells.
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