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U2OS E2F7 KO cell line

Invented by Ana Maria Zubiaga, at University of the Basque Country, EHU Iraia Garcia Santisteban at University of the Basque Country, EHU

Info

Catalogue Number 156469
Antigen/Gene or Protein Targets Full knockout for the E2F7 gene
Parental Line U2OS
Synonyms U-2 OS
Host Human
Tissue Bone
Disease Keywords Osteosarcoma cell line
Model Knock-Out
Relevance 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 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.
Conditional No
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

References

There are 1 reference entries for this reagent.

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References: 1 entry

Mitxelena et al. 2018. Nucleic Acids Res. 46(9):4546-4559. PMID: 29590434.


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References: 1 entry

Mitxelena et al. 2018. Nucleic Acids Res. 46(9):4546-4559. PMID: 29590434.


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Inventor Information

Inventors

Ana Maria Zubiaga

Ana Maria Zubiaga

Iraia Garcia Santisteban

Iraia Garcia Santisteban

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