- Name Ana Maria Zubiaga ; Iraia Garcia Santisteban
- Institute University of the Basque Country (EHU)
- Tool name: U2OS E2F7 KO cell line
- Alternate names: U-2 OS
- Tool type: Cell Lines
- Tool sub-type: Continuous
- Parental cell line: U2OS
- Organism: Human
- Tissue: Bone
- Cancer type: Osteosarcoma cell line
- Disease: Cancer
- Growth properties: Increased G2/M checkpoint recovery competence and improved viability upon treatment with compounds that affect replication fork progression (cisplatin, mitomycin C, olaparib).
- Model: Knock-Out
- Conditional: No
- Description: E2F7 is a transcription factor that participates in various processes such as cell cycle regulation and DNA damage response by mediating transcriptional repression of target genes involved in G1/S regulation as well as in DNA replication and repair. E2F7 behaves as a tumour suppressor by inhibiting DNA repair activity of tumour cells upon chemotherapy treatment. Loss of E2F7 by CRISPR knockout confers increased resistance to chemotherapy in BRCA2-deficient cells. CRISPR edited U2OS cells.
- Research area: Cancer; Cell Cycle; Other
- Production details: E2F7 knockout cells were generated using the CRISPR/Cas9 system. A CRISPR guide RNA (gRNA) targeting the first coding exon of E2F7 was designed using Benchling, and cloned into the BbsI site of pX330 (42230, Addgene). U2OS cells were co-transfected with this plasmid, together with a plasmid containing a gRNA to the zebrafish TIA gene (5 -GGTATGTCGGGAACCTCTCC3 ) and a P2A-puromycin resistance cassette flanked by two TIA target sites. Co-transfection results in excision of the cassette and subsequent sporadic incorporation at the site of the targeted genomic locus. Successful integration of the cassette into the targeted gene disrupts the allele and renders cells resistant to puromycin. After puromycin selection, resistant clones were expanded, screened for cassette integration and indels into the target gene.
- Additional notes: CRISPR edited U2OS cells. Cancer Research Technology Limited (trading research tools as Ximbio) has been granted a non-exclusive license to the CRISPR-Cas9 technology by ERS Genomics Ltd under the patent rights listed here. This license from ERS Genomics Ltd allows Ximbio to develop and commercialise CRISPR-Cas9 modified cell lines for research use only. Ximbio can provide these modified CRISPR-Cas9 cell lines to companies under a label-use only license.
- For Research Use Only
- Target: Full knockout for the E2F7 gene
- Application notes: Cancer Research Technology Limited (trading research tools as CancerTools.org) has been granted a non-exclusive license to the CRISPR-Cas9 technology by ERS Genomics Ltd under the patent rights listed here: https://www.cancertools.org/tool-faqs#hs_cos_wrapper_widget_1649861453796 This license from ERS Genomics Ltd allows CancerTools.org to develop and commercialise CRISPR-Cas9 modified cell lines for research use only. CancerTools.org can provide these modified CRISPR-Cas9 cell lines to companies under a label-use only license
- Format: Frozen
- Growth medium: Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS)
- Storage conditions: Liquid Nitrogen
- Shipping conditions: Dry ice
- Mycoplasma free: Yes
- Biosafety level: 1
- • Mitxelena et al. 2018. Nucleic Acids Res. 46(9):4546-4559. PMID: 29590434.