Human DMPK Knockout HCT116 Cell Line

Storage Condition	Upon arrival, the vial should be stored in liquid nitrogen vapor phase. Storage buffer: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether
Shipping Condition	Shipped on dry ice.

Parental Cell Line	HCT116
Passage Number	<20
Viability	~80%
Mutation Description	CRISPR/Cas9 system is used to generate knockout cell lines. Homozygous: 1 bp insertion in exon 1.
Adherent / Suspension	Adherent
Cell Type	Epithelial Cells
Mycoplasma Free	Yes
Knockout Validation	Sanger Sequencing
Cell Medium	Culture medium: McCoY5a + 10% FBS. Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Initial Handling Guidelines	1. Thaw the vial in 37ºC water bath approximately 1-2 minutes. 2. Transfer the cell suspension to a 15 ml/50 ml conical sterile polypropylene centrifuge tube containing pre-warmed culture medium. Wash vial with an additional culture medium to collect remaining cells, and centrifuge at 201 × g (rcf) for 5 minutes at room temperature. 3. Resuspend the cell pellet in 5 ml pre-warmed culture medium and count cell number. Seed cells in an appropriate cell culture flask at a density of 2×10⁴ cells/cm². This should allow for confluency within 48 hours. 4. Incubate the culture at 37ºC incubator with 5% CO₂. Cultures should be monitored daily.
Subculture Guidelines	Cells should be passaged when they have achieved 80-90% confluence. A partial media change 24 hours prior to subculture may be helpful to encourage growth.
Disease	Carcinoma
Gender	Male
Description	Human wild-type HCT116 cell line as the recommended control.

Tissue Specificity	Most isoforms are expressed in many tissues, including brain, heart, liver, and skeletal muscle. Isoform 2 is only found in the heart and skeletal muscle. Isoform 14 is only found in the brain.
Target Sequence Similarities	Belongs to the AGC Ser/Thr protein kinase family, DMPK subfamily.