ATM Knockout cell line(HCT116)

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ATM Knockout cell line(HCT116)

ATM Knockout cell line(HCT116) Order

Catalog#YKO-H600

Price (USD)-

Size 1*10^6

Instruction

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

Cell Info

Catalog YKO-H600
Cell line ATM Knockout cell line(HCT116) Cell Type Human Colon Cancer Cell Line
Morphology Epithelial-like, adherent Passage ratio 1:3~1:4
Culture method 90%McCOY's 5A+10% FBS
Cryopreservation solution 50%McCOY's 5A+40% FBS+10%DMSO
Special Note

STR Validation

Gene Information

Official symbol
ATM
Gene id
472
Official full symbol
ATM serine/threonine kinase
Also known as
AT1, ATA, ATC, ATD, ATDC, ATE, TEL1, TELO1
The protein encoded by this gene belongs to the PI3/PI4-kinase family. This protein is an important cell cycle checkpoint kinase that phosphorylates; thus, it functions as a regulator of a wide variety of downstream proteins, including tumor suppressor proteins p53 and BRCA1, checkpoint kinase CHK2, checkpoint proteins RAD17 and RAD9, and DNA repair protein NBS1. This protein and the closely related kinase ATR are thought to be master controllers of cell cycle checkpoint signaling pathways that are required for cell response to DNA damage and for genome stability. Mutations in this gene are associated with ataxia telangiectasia, an autosomal recessive disorder.

KO strategy

Knockout region: Exon 11 (based on transcript ATM-229)
Validation: PCR+Sanger Sequencing

Method

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

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