OTULIN Knockout cell line (HepG2)

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OTULIN Knockout cell line (HepG2)

OTULIN Knockout cell line (HepG2) Order

Catalog#YKO-H784

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-H784
Cell line OTULIN Knockout cell line (HepG2) Cell Type Human Hepatoma Cell Line
Morphology Epithelial-like, adherent Passage ratio 1:3~1:5
Culture method 90% DMEM+10% FBS
Cryopreservation solution 50%DMEM+40%FBS+10%DMSO
Special Note

STR Validation

Gene Information

Official symbol
OTULIN
Gene id
90268
Official full symbol
OTU deubiquitinase with linear linkage specificity
Also known as
AIPDS, FAM105B, GUM
This gene encodes a member of the peptidase C65 family of ubiquitin isopeptidases. Members of this family remove ubiquitin from proteins. The encoded enzyme specifically recognizes and removes M1(Met1)-linked, or linear, ubiquitin chains from protein substrates. Linear ubiquitin chains are known to regulate the NF-kappa B signaling pathway in the context of immunity and inflammation. Mutations in this gene cause a potentially fatal autoinflammatory syndrome in human patients.

KO strategy

Knockout region: Exon 3 (based on transcript OTULIN-201)
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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