ArticleScientific reports2018
CRISPR/Cas9 mediated targeting of multiple genes in Dictyostelium.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 60 papers.
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Who cites it
60 citing papers in PubMed.
- Unlocking CRISPR-Cas9 editing for widely diverse Dictyostelid species.Molecular systems biology · 2026Article
- Early cAMP signaling orchestrates single-cell synchronicity throughout Dictyostelium development.Communications biology · 2026Article
- A nanobody-based degron system for targeted protein knockdown in Dictyostelium discoideum.BMC molecular and cell biology · 2026Article
- Genome editing across Dictyostelia species enables comparative functional genetics of social amoebas.Scientific reports · 2026Article
- The RNA-binding protein RNP1A is essential and interacts with contractility kit proteins to facilitate cell mechanics.Journal of cell science · 2026Article
- Protein arginine methyltransferases in protozoan parasites: a new path for antiparasitic chemotherapy?Microbial cell (Graz, Austria) · 2026Article
- Global profiling of polyketide synthases in facultative multicellular eukaryotes.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Cells adapt to extracellular acidic pH through TM9SF3-mediated PI(4,5)PNature communications · 2025Article
- Luminal Phospholipase D Attacks Bacterial Membranes in Dictyostelium discoideum Phagosomes.Molecular microbiology · 2025Article
- Excitable Ras dynamics-based screens reveal RasGEFX is required for macropinocytosis and random cell migration.Nature communications · 2025Article
- Efficient endogenous protein labelling in Dictyostelium using CRISPR/Cas9 knock-in and split fluorescent proteins.PloS one · 2025Article
- The Amoeba Dictyostelium discoideum as Novel Production Host for Complex Substances.Advances in biochemical engineering/biotechnology · 2025Review
- Agrobacterium-Mediated Transformation and Targeted Mutagenesis Using SpCas12f in Rice.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Optogenetic control of cAMP oscillations reveals frequency-selective transcription factor dynamics in Dictyostelium.Development (Cambridge, England) · 2025Article
- Antibacterial effectors inmSphere · 2024Article
- mTORC1 pathway activity biases cell fate choice.Scientific reports · 2024Article
- Pharmacological inhibition of ENT1 enhances the impact of specific dietary fats on energy metabolism gene expression.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- RapB Regulates Cell Adhesion and Migration in Dictyostelium, Similar to RapA.Journal of microbiology (Seoul, Korea) · 2024Article
- Developing a Tanshinone IIA Memetic by Targeting MIOS to Regulate mTORC1 and Autophagy in Glioblastoma.International journal of molecular sciences · 2024Article
- Functional mechanism study of the allelochemical myrigalone A identifies a group of ultrapotent inhibitors of ethylene biosynthesis in plants.Plant communications · 2024Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR/Cas9 has emerged in various organisms as a powerful technology for targeted gene knockout; however, no reports of editing the Dictyostelium genome efficiently using this system are available. We describe here the application of CRISPR/Cas9-mediated gene modification in Dictyostelium. The endogenous tRNA-processing system for expressing sgRNA was approximately 10 times more effective than the commonly used U6 promoter. The resulting sgRNA affected the sub-nuclear localisation of Cas9, indicating that the expression level of sgRNA was sufficiently high to form Cas9 and sgRNA complexes within the nucleus. The all-in-one vector containing Cas9 and sgRNA was transiently expressed to generate mutants in five PI3K genes. Mutation detective PCR revealed the mutagenesis frequency of the individual genes to be between 72.9% and 100%. We confirmed that all five targeting loci in the four independent clones had insertion/deletion mutations in their target sites. Thus, we show that the CRISPR/Cas9 system can be used in Dictyostelium cells to enable efficient genome editing of multiple genes. Since this system utilises transient expression of the all-in-one vector, it has the advantage that the drug resistance cassette is not integrated into the genome and simple vector construction, involving annealing two oligo-DNAs.
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