ArticleScientific reports2020
A simple method using CRISPR-Cas9 to knock-out genes in murine cancerous cell lines.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed, 32 citations in OpenAlex.
- CROP: a feature-independent context-aware method for CRISPR-Cas9 frameshift prediction.Bioinformatics (Oxford, England) · 2026Article
- Biological mechanism and immune response of MHC-II expression in tumor cells.Cancer biology & medicine · 2025Review
- HVJ-E links Apolipoprotein d to antitumor effects.Journal for immunotherapy of cancer · 2025Article
- Translating time: Challenges, progress, and future directions.Brain research bulletin · 2025Review
- BioID-based intact cell interactome of the Kv1.3 potassium channel identifies a Kv1.3-STAT3-p53 cellular signaling pathway.Science advances · 2024Article
- Streamlined Generation of CRISPR/Cas9-Mediated Single-Cell Knockout Clones in Murine Cell Lines.ACS pharmacology & translational science · 2024Article
- Comparison of antibody-based immunotherapeutics for malignant hematological disease in an experimental murine model.Blood advances · 2024Article
- Molecular and Spatial Signatures of Mouse Embryonic Endothelial Cells at Single-Cell Resolution.Circulation research · 2024Article
- The two-faced role of RNA methyltransferase METTL3 on cellular response to cisplatin in head and neck squamous cell carcinomaFrontiers in oncology · 2024Article
- CRISPR/Cas9 systems: Delivery technologies and biomedical applications.Asian journal of pharmaceutical sciences · 2023Review
- IL2-inducible T-cell Kinase is Required for HBV-induced Type T Interferon Expression and Antiviral Response.Journal of clinical and translational hepatology · 2023Article
- New CRISPR Technology for Creating Cell Models of Lipoprotein Assembly and Secretion.Current atherosclerosis reports · 2023Review
- Molecular and therapeutic effect of CRISPR in treating cancer.Medical oncology (Northwood, London, England) · 2023Review
- N-Acetylglucosamine Sensing and Metabolic Engineering for Attenuating Human and Plant Pathogens.Bioengineering (Basel, Switzerland) · 2022Review
- Fast-Track and Integration-Free Method of Genome Editing by CRISPR/Cas9 in Murine Pluripotent Stem Cells.Frontiers in cell and developmental biology · 2022Article
- Function of cofactor Akirin2 in the regulation of gene expression in model human Caucasian neutrophil-like HL60 cells.Bioscience reports · 2021Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR-Cas9 system can be used to generate knock-out cancer cell lines. An insertion or deletion induced by a single guide RNA (gRNA) is often used to generate knock-out cells, however, some cells express the target gene by skipping the disrupted exon, or by using a splicing variant, thus losing the target exon. To overcome this unexpected expression of the target gene, almost the entire gene can be swapped with a selection marker. However, it is time-consuming to create a targeting vector which contains 5' and 3' homology arms flanked by a selection marker. Here, we developed a simple and easy method called SUCCESS (Single-strand oligodeoxynucleotides, Universal Cassette, and CRISPR/Cas9 produce Easy Simple knock-out System), to knock-out a target gene without constructing a targeting vector. Our method removed the targeted large genomic region by using two pX330 plasmids encoding Cas9 and gRNA, two 80mer single strand oligodeoxynucleotides (ssODN), and a blunt-ended universal selection maker sequence in B16F10 murine cancer cell and ID8 murine ovarian cancer cell. SUCCESS generated knock-out clones in two murine cancer cell lines by homozygous deletion of the target genomic region, and without constructing targeting vectors. Thus, our method can be widely applied to generate homozygous knock-out cell lines, as well as knock-in cell lines.
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