ArticleNucleic acids research2020
Synthetic CRISPR/Cas9 reagents facilitate genome editing and homology directed repair.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 48 citations in OpenAlex.
- Optimised genome editing for precise DNA insertion and substitution using prime editors in zebrafish.eLife · 2026Article
- Genetically encoded affinity reagents are a toolkit for visualizing and manipulating endogenous protein function in vivo.Nature communications · 2025Article
- Identifying optimal conditions for precise knock-in of exogenous DNA into the zebrafish genome.Development (Cambridge, England) · 2025Article
- Zebrafishology, study design guidelines for rigorous and reproducible data using zebrafish.Communications biology · 2025Review
- The piRNA protein Asz1 is essential for germ cell and gonad development in zebrafish and exhibits differential necessities in distinct types of germ granules.PLoS genetics · 2025Article
- Instantaneous visual genotyping and facile site-specific transgenesis via CRISPR-Cas9 and phiC31 integrase.Biology open · 2024Article
- Melanocytes in regenerative medicine applications and disease modeling.Journal of translational medicine · 2024Review
- Article
- Efficient knock-in method enabling lineage tracing in zebrafish.Life science alliance · 2023Article
- Review
- New advances in CRISPR/Cas-mediated precise gene-editing techniques.Disease models & mechanisms · 2023Review
- In search of an ideal template for therapeutic genome editing: A review of current developments for structure optimization.Frontiers in genome editing · 2023Review
- Activation of a transient progenitor state in the epicardium is required for zebrafish heart regeneration.Nature communications · 2022Article
- A robust pipeline for efficient knock-in of point mutations and epitope tags in zebrafish using fluorescent PCR based screening.BMC genomics · 2022Article
- SpG and SpRY variants expand the CRISPR toolbox for genome editing in zebrafish.Nature communications · 2022Article
- Generation of a zebrafish knock-in line expressing MYC-tagged Sox11a using CRISPR/Cas9 genome editing.Biochemical and biophysical research communications · 2022Article
- Review
- Key sequence features of CRISPR RNA for dual-guide CRISPR-Cas9 ribonucleoprotein complexes assembled with wild-type or HiFi Cas9.Nucleic acids research · 2022Article
- Pigment pattern morphospace of Danio fishes: evolutionary diversification and mutational effects.Biology open · 2021Article
- Progress in Gene-Editing Technology of Zebrafish.Biomolecules · 2021Review
Corrections and comments
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Authors and funding
16 authors at 3 institutions in 1 country.
Funding
Abstract
CRISPR/Cas9 has become a powerful tool for genome editing in zebrafish that permits the rapid generation of loss of function mutations and the knock-in of specific alleles using DNA templates and homology directed repair (HDR). We examined the efficiency of synthetic, chemically modified gRNAs and demonstrate induction of indels and large genomic deletions in combination with recombinant Cas9 protein. We developed an in vivo genetic assay to measure HDR efficiency and we utilized this assay to test the effect of altering template design on HDR. Utilizing synthetic gRNAs and linear dsDNA templates, we successfully performed knock-in of fluorophores at multiple genomic loci and demonstrate transmission through the germline at high efficiency. We demonstrate that synthetic HDR templates can be used to knock-in bacterial nitroreductase (ntr) to facilitate lineage ablation of specific cell types. Collectively, our data demonstrate the utility of combining synthetic gRNAs and dsDNA templates to perform homology directed repair and genome editing in vivo.
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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.