ArticleNature protocols2014
CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish.
Article in Nature protocols, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- Dynamic interactions between epithelial skin cells and a sensory cavity sculpt the growing olfactory orifice.Nature communications · 2026Article
- Pioneer statoacoustic neurons guide neuroblast behaviour during otic ganglion assembly.Development (Cambridge, England) · 2023Article
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- Actomyosin contractility in olfactory placode neurons opens the skin epithelium to form the zebrafish nostril.Developmental cell · 2023Article
- A transgenic system for targeted ablation of reproductive and maternal-effect genes.Development (Cambridge, England) · 2021Article
- Expression of a Barhl1a reporter in subsets of retinal ganglion cells and commissural neurons of the developing zebrafish brain.Scientific reports · 2020Article
- Actomyosin regulation by Eph receptor signaling couples boundary cell formation to border sharpness.eLife · 2019Article
- Article
- Functional analysis of the UL24 protein of suid herpesvirus 1.Virus genes · 2019Article
- Using CRISPR/Cas9 for Gene Knockout in Immunodeficient NSG Mice.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Application of CRISPR/Cas9-Mediated Gene Editing in Tomato.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Generation and characterization of UL41 null pseudorabies virus variant in vitro and in vivo.Virology journal · 2018Article
- Alk and Ltk ligands are essential for iridophore development in zebrafish mediated by the receptor tyrosine kinase Ltk.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Opportunities for CRISPR/Cas9 Gene Editing in Retinal Regeneration Research.Frontiers in cell and developmental biology · 2017Review
- A simplified method for identifying early CRISPR-induced indels in zebrafish embryos using High Resolution Melting analysis.BMC genomics · 2016Article
- Advances and perspectives in the application of CRISPR/Cas9 in insects.Dong wu xue yan jiu = Zoological research · 2016Review
- 2C-Cas9: a versatile tool for clonal analysis of gene function.Genome research · 2016Article
- Targeted isolation and cloning of 100-kb microbial genomic sequences by Cas9-assisted targeting of chromosome segments.Nature protocols · 2016Article
- Tracking Cells in GFP-transgenic Zebrafish Using the Photoconvertible PSmOrange System.Journal of visualized experiments : JoVE · 2016Article
- ssODN-mediated knock-in with CRISPR-Cas for large genomic regions in zygotes.Nature communications · 2016Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Here we present a protocol for the conversion of eGFP-transgenic zebrafish lines into lines expressing Gal4 from the same locus. This conversion allows the in-depth analysis of the former eGFP-expressing cell population; with the Gal4-upstream activating sequence (UAS) system, diverse UAS transgenes can be transactivated. Site-specific targeting of the gene encoding eGFP is achieved using the clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) system. A single-guide RNA (sgRNA) that targets eGFP is injected into embryos together with a donor vector containing an optimized version of Gal4 (KalTA4) to trigger integration of the donor into the targeted eGFP genomic location. To enable screening for successful integration events, injection is performed in a UAS:RFP transgenic background; fish showing mosaic eGFP-to-RFP conversion are raised to adulthood. The progeny of these adult fish are then screened for stable germline transmission, and converted progeny are used to generate stable lines. We have been able to generate two stably converted transgenic lines within 4 months.
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