ArticleGenetics2016
SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification in Caenorhabditis elegans.
Article in Genetics, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 130 papers.
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Who cites it
130 citing papers in PubMed.
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- SynaptoTagMe, a toolkit for in vivo mapping and modulating neurotransmission at single-cell resolution.eLife · 2026Article
- Tissue-specific DNA repair strategies underlie apparent high transposon activity in the Caenorhabditis elegans soma.G3 (Bethesda, Md.) · 2026Article
- Impact of Fluorophore and Epitope Position on Destabilized Reporter Performance inbioRxiv : the preprint server for biology · 2026Article
- Engineering the Caenorhabditis elegans genome with a nested, self-excising selection cassette.Genetics · 2026Article
- Katanin-mediated severing generates microtubules during neurite outgrowth.Research square · 2026Article
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- Roles and regulation of the Kunitz domain protein MLT-11 during C. elegans cuticle synthesis and molting.Genetics · 2026Article
- TAG-IN, a swappable strategy for endogenous gene tagging inbioRxiv : the preprint server for biology · 2026Article
- Modular gene tagging inbioRxiv : the preprint server for biology · 2026Article
- An F2A sequence permits correct localization of a secreted and a nuclear localized reporter inmicroPublication biology · 2026Article
- Article
- MLT-11 is necessary forbioRxiv : the preprint server for biology · 2025Article
- Transposon Regulation in thebioRxiv : the preprint server for biology · 2025Article
- AnalysisbioRxiv : the preprint server for biology · 2025Article
- Expansion of lysosomal capacity in early adult neurons driven by TFEB/HLH-30 protects dendrite maintenance during aging in Caenorhabditis elegans.PLoS biology · 2025Article
- Multiscale patterning of a model apical extracellular matrix revealed by systematic endogenous protein tagging.bioRxiv : the preprint server for biology · 2025Article
- Rab GTPases are evolutionarily conserved signals mediating selective autophagy.The Journal of cell biology · 2025Article
- A modular system to label endogenous presynaptic proteins using split fluorophores in Caenorhabditis elegans.Genetics · 2025Article
70 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors.
Funding
Abstract
In principle, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 allows genetic tags to be inserted at any locus. However, throughput is limited by the laborious construction of repair templates and guide RNA constructs and by the identification of modified strains. We have developed a reagent toolkit and plasmid assembly pipeline, called "SapTrap," that streamlines the production of targeting vectors for tag insertion, as well as the selection of modified Caenorhabditis elegans strains. SapTrap is a high-efficiency modular plasmid assembly pipeline that produces single plasmid targeting vectors, each of which encodes both a guide RNA transcript and a repair template for a particular tagging event. The plasmid is generated in a single tube by cutting modular components with the restriction enzyme SapI, which are then "trapped" in a fixed order by ligation to generate the targeting vector. A library of donor plasmids supplies a variety of protein tags, a selectable marker, and regulatory sequences that allow cell-specific tagging at either the N or the C termini. All site-specific sequences, such as guide RNA targeting sequences and homology arms, are supplied as annealed synthetic oligonucleotides, eliminating the need for PCR or molecular cloning during plasmid assembly. Each tag includes an embedded Cbr-unc-119 selectable marker that is positioned to allow concurrent expression of both the tag and the marker. We demonstrate that SapTrap targeting vectors direct insertion of 3- to 4-kb tags at six different loci in 10-37% of injected animals. Thus SapTrap vectors introduce the possibility for high-throughput generation of CRISPR/Cas9 genome modifications.
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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.