ArticleGenome research2014
Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair.
Article in Genome research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 323 papers.
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Who cites it
323 citing papers in PubMed, 646 citations in OpenAlex.
- Interfering with DNA repair pathways to enhance CRISPR-Cas9-mediated homology-directed repair in a chelicerate genetic model.iScience · 2026Article
- Catfish Transgenesis and Gene Editing: Overview, Recent Advances, and Future Perspectives.Biology · 2026Review
- A Reproducible Electroporation Strategy for CRISPR-Cas9 RNP and mRNA Delivery in Fish Embryos.Marine biotechnology (New York, N.Y.) · 2026Article
- Review
- Tunable gene expression in zebrafish using RiboSCALE.bioRxiv : the preprint server for biology · 2026Article
- Versatile CRISPR-Cas Tools for Gene Regulation in Zebrafish via an Enhanced Q Binary System.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Advancing knock-in approaches for robust genome editing in zebrafish.Biology open · 2026Article
- Cell sources and engineering strategies for cultured meat production: a critical comparative review.Frontiers in nutrition · 2026Review
- Dynamic junction remodeling and transient vascular disruption during cancer cell extravasation across the blood-brain barrier.Frontiers in cell and developmental biology · 2026Article
- Advancing Knock-In Approaches for Robust Genome Editing in Zebrafish.bioRxiv : the preprint server for biology · 2025Article
- Article
- Genetic requirement for Esrp1 and Esrp2 in vertebrate pituitary morphogenesis.Development (Cambridge, England) · 2025Article
- Jam2 Signaling Functions Downstream of Hand2 To Initiate The Formation Of Organ-Specific Vascular Progenitors In Zebrafish.bioRxiv : the preprint server for biology · 2025Article
- Gene and Stem Cell-Based Therapies for Retinal Degenerative Diseases: Update, Challenges, and Future Directions.Stem cell reviews and reports · 2025Review
- Advances in large-scale DNA engineering with the CRISPR system.Experimental & molecular medicine · 2025Review
- Non-viral intron knock-ins for targeted gene integration into human T cells and for T-cell selection.Nature biomedical engineering · 2025Article
- Zebrafish Models of Induced Lymphangiogenesis: Current Advancements and Therapeutic Discovery.Pharmaceuticals (Basel, Switzerland) · 2025Review
- Pu.1/Spi1 dosage controls the turnover and maintenance of microglia in zebrafish and mammals.eLife · 2025Article
- Transferable approaches to CRISPR-Cas9 induced genome editing in non-model insects: a brief guide.Frontiers in zoology · 2025Review
- Cloning-Free Targeting of Endogenous Loci to Generate Fluorescent Reporters in Medaka.Bio-protocol · 2025Article
263 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Sequence-specific nucleases like TALENs and the CRISPR/Cas9 system have greatly expanded the genome editing possibilities in model organisms such as zebrafish. Both systems have recently been used to create knock-out alleles with great efficiency, and TALENs have also been successfully employed in knock-in of DNA cassettes at defined loci via homologous recombination (HR). Here we report CRISPR/Cas9-mediated knock-in of DNA cassettes into the zebrafish genome at a very high rate by homology-independent double-strand break (DSB) repair pathways. After co-injection of a donor plasmid with a short guide RNA (sgRNA) and Cas9 nuclease mRNA, concurrent cleavage of donor plasmid DNA and the selected chromosomal integration site resulted in efficient targeted integration of donor DNA. We successfully employed this approach to convert eGFP into Gal4 transgenic lines, and the same plasmids and sgRNAs can be applied in any species where eGFP lines were generated as part of enhancer and gene trap screens. In addition, we show the possibility of easily targeting DNA integration at endogenous loci, thus greatly facilitating the creation of reporter and loss-of-function alleles. Due to its simplicity, flexibility, and very high efficiency, our method greatly expands the repertoire for genome editing in zebrafish and can be readily adapted to many other organisms.
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Registered trials
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.