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- Inventor Info
|Antigen/Gene or Protein Targets||Full knockout for the E2F7 gene|
|Disease Keywords||Osteosarcoma cell line|
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.
|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.|
|Research Area||Cancer, Cell Cycle, Other|
|Growth/Phenotype Keywords||Increased G2/M checkpoint recovery competence and improved viability upon treatment with compounds that affect replication fork progression (cisplatin, mitomycin C, olaparib).|
|Recommended Growing Conditions||Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS)|
|Positive Control||Parental U2OS cells, puromycin sensitive|
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.
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Ana Maria Zubiaga
Iraia Garcia Santisteban