JAK3 Knockout cell line (Jurkat)

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JAK3 Knockout cell line (Jurkat)

JAK3 Knockout cell line (Jurkat) Order

Catalog#YKO-H763

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-H763
Cell line JAK3 Knockout cell line(Jurkat, Clone E6-1) Cell Type Human T lymphocyte cell line
Morphology Lymphocyte-like, 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 50%RPMI-1640+40%FBS+10%DMSO
Special Note The cell culture density needs to be strictly controlled.

STR Validation

Gene Information

Official symbol
JAK3
Gene id
3718
Official full symbol
Janus kinase 3
Also known as
JAK-3, JAK3_HUMAN, JAKL, L-JAK, LJAK
The protein encoded by this gene is a member of the Janus kinase (JAK) family of tyrosine kinases involved in cytokine receptor-mediated intracellular signal transduction. It is predominantly expressed in immune cells and transduces a signal in response to its activation via tyrosine phosphorylation by interleukin receptors. Mutations in this gene are associated with autosomal SCID (severe combined immunodeficiency disease).

KO strategy

Knockout region: Exon 2 (based on transcript JAK3-201)
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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