VPS4B Knockout cell line (HCT 116)

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VPS4B Knockout cell line (HCT 116)

VPS4B Knockout cell line (HCT 116) Order

Catalog#YKO-H673

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-H673
Cell line VPS4B 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
VPS4B
Gene id
9525
Official full symbol
vacuolar protein sorting 4 homolog B
Also known as
MIG1, SKD1, SKD1B, VPS4-2
The protein encoded by this gene is a member of the AAA protein family (ATPases associated with diverse cellular activities), and is the homolog of the yeast Vps4 protein. In humans, two paralogs of the yeast protein have been identified. The former share a high degree of aa sequence similarity with each other, and also with yeast Vps4 and mouse Skd1 proteins. Mouse Skd1 (suppressor of K+ transport defect 1) has been shown to be a yeast Vps4 ortholog. Functional studies indicate that both human paralogs associate with the endosomal compartments, and are involved in intracellular protein trafficking, similar to Vps4 protein in yeast. The gene encoding this paralog has been mapped to chromosome 18; the gene for the other resides on chromosome 16.

KO strategy

Knockout region: Exon 3 (based on transcript VPS4B-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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