PRAME Knockout cell line (HCT116)

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PRAME Knockout cell line (HCT116)

PRAME Knockout cell line (HCT116) Order

Catalog#YKO-H723

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-H723
Cell line PRAME Knockout cell line(HCT 116) 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
PRAME
Gene id
23532
Official full symbol
PRAME nuclear receptor transcriptional regulator
Also known as
CT130, MAPE, OIP-4, OIP4
This gene encodes an antigen that is preferentially expressed in human melanomas and that is recognized by cytolytic T lymphocytes. It is not expressed in normal tissues, except testis. The encoded protein acts as a repressor of retinoic acid receptor, and likely confers a growth advantage to cancer cells via this function. Alternative splicing results in multiple transcript variants.

KO strategy

Knockout region: Exon 4 (based on transcript PRAME-205)
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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