ArticleFrontiers in cell and developmental biology2021
Highly Efficient Synthetic CRISPR RNA/Cas9-Based Mutagenesis for Rapid Cardiovascular Phenotypic Screening in F0 Zebrafish.
Article in Frontiers in cell and developmental biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 27 citations in OpenAlex.
- Review
- Efficient Gene Expression System in Medaka Embryos Enables Functional Characterization of nt5c1a Paralogs Involved in Inosine Monophosphate Metabolism.Genesis (New York, N.Y. : 2000) · 2026Article
- Single-Cell-Resolution Fate Mapping Reveals Embryonic Venous Origins of Fenestrated Hindbrain Choroid Plexus Vasculature.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2026Article
- Review
- Zebrafish as a model for Catel-Manzke syndrome-identification and characterization of the zebrafish TGDS ortholog.The FEBS journal · 2026Article
- Bnip3lb-driven mitophagy maintains fate of the embryonic hematopoietic stem cell pool.Nature communications · 2026Article
- Apelin signaling acts as a molecular switch between endothelial and hematopoietic stem cell fates.EMBO reports · 2026Article
- CRISPR/Cas9-Based Mutagenesis Strategies for Efficient Biallelic Gene Inactivation and Consistent Phenotypic Detection in F0 Zebrafish.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Zebrafish: A Versatile and Powerful Model for Biomedical Research.BioEssays : news and reviews in molecular, cellular and developmental biology · 2025Review
- Further delineation of defects in MRPS2 causing human OXPHOS deficiency and early developmental abnormalities in zebrafish.European journal of human genetics : EJHG · 2025Article
- In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor.Nature communications · 2025Article
- A Rapid F0 CRISPR Screen in Zebrafish to Identify Regulator Genes of Neuronal Development in the Enteric Nervous System.Neurogastroenterology and motility · 2025Article
- Optimizing gRNA selection for high-penetrance F0 CRISPR screening for interrogating disease gene function.Nucleic acids research · 2025Article
- Neural progenitor-derived Apelin controls tip cell behavior and vascular patterning.Science advances · 2024Article
- Genomic screening of fish-specific genes in gnathostomes and their functions in fin development.Development, growth & differentiation · 2024Article
- Large-scale F0 CRISPR screens in vivo using MIC-Drop.Nature protocols · 2023Review
- Article
- Gene Modulation with CRISPR-based Tools in Human iPSC-Cardiomyocytes.Stem cell reviews and reports · 2023Review
- Aquatic Freshwater Vertebrate Models of Epilepsy Pathology: Past Discoveries and Future Directions for Therapeutic Discovery.International journal of molecular sciences · 2022Review
- Historical and current perspectives on blood endothelial cell heterogeneity in the brain.Cellular and molecular life sciences : CMLS · 2022Review
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
The zebrafish is a valuable vertebrate model to study cardiovascular formation and function due to the facile visualization and rapid development of the circulatory system in its externally growing embryos. Despite having distinct advantages, zebrafish have paralogs of many important genes, making reverse genetics approaches inefficient since generating animals bearing multiple gene mutations requires substantial efforts. Here, we present a simple and robust synthetic CRISPR RNA/Cas9-based mutagenesis approach for generating biallelic F0 zebrafish knockouts. Using a dual-guide synthetic CRISPR RNA/Cas9 ribonucleoprotein (dgRNP) system, we compared the efficiency of biallelic gene disruptions following the injections of one, two, and three dgRNPs per gene into the cytoplasm or yolk. We show that simultaneous cytoplasmic injections of three distinct dgRNPs per gene into one-cell stage embryos resulted in the most efficient and consistent biallelic gene disruptions. Importantly, this triple dgRNP approach enables efficient inactivation of cell autonomous and cell non-autonomous gene function, likely due to the low mosaicism of biallelic disruptions. In support of this finding, we provide evidence that the F0 animals generated by this method fully phenocopied the endothelial and peri-vascular defects observed in corresponding stable mutant homozygotes. Moreover, this approach faithfully recapitulated the trunk vessel phenotypes resulting from the genetic interaction between two
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.