NR1H4 Knockout cell line (Huh-7)

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NR1H4 Knockout cell line (Huh-7)

NR1H4 Knockout cell line (Huh-7) Order

Catalog#YKO-H660

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-H660
Cell line NR1H4 Knockout cell line(HuH-7) Cell Type Human Hepatoma Cell Line
Morphology Epithelial-like, adherent Passage ratio 1:2~1:3
Culture method 90%DMEM+10% FBS
Cryopreservation solution 50%DMEM+40% FBS+10%DMSO
Special Note

STR Validation

Gene Information

Official symbol
NR1H4
Gene id
9971
Official full symbol
nuclear receptor subfamily 1 group H member 4
Also known as
BAR, FXR, HRR-1, HRR1, PFIC5, RIP14
This gene encodes a ligand-activated transcription factor that shares structural features in common with nuclear hormone receptor family members. This protein functions as a receptor for bile acids, and when bound to bile acids, binds to DNA and regulates the expression of genes involved in bile acid synthesis and transport. Alternatively spliced transcript variants encoding different isoforms have been described.

KO strategy

Knockout region: Exon 4 (based on transcript NR1H4-203)
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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