GSDMD Knockout cell line(THP-1)

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GSDMD Knockout cell line(THP-1)

GSDMD Knockout cell line(THP-1) Order

Catalog#YKO-H172

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-H172
Cell line GSDMD Knockout cell line(THP-1) Cell Type Human Ovarian Granulosa Cell Line
Morphology Monocyte, suspension Passage ratio Control the cell density at 2x10^5~4.0x10^5 cells/ml during passaging, and carry out the passage when the cells grow to 8x10^5~1.0x10^6 cells/ml.
Culture method 90%RPMI-1640+10%FBS
Cryopreservation solution 90%FBS+10%DMSO
Special Note The cell culture density needs to be strictly controlled. The cell viability after thawing is relatively low, it is recommended to use Australia-sourced FBS for thawing or increase the FBS ratio to 20%. When the cells grow normally and can be passaged after 2 passages, the cells can be cultured using the normal culture method.

STR Validation

Gene Information

Official symbol
GSDMD
Gene id
79792
Official full symbol
gasdermin D
Also known as
DF5L, DFNA5L, FKSG10, GSDMDC1
Gasdermin D is a member of the gasdermin family. Members of this family appear to play a role in regulation of epithelial proliferation. Gasdermin D has been suggested to act as a tumor suppressor. Alternatively spliced transcript variants have been described.

KO strategy

Knockout region: Exon 2 (based on transcript GSDMD-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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