KC-0875

293T-SLC39A6-KO-3B3-Cell-Line

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Background of 293T-SLC39A6-KO-3B3-Cell-Line

Zinc is an essential cofactor for hundreds of enzymes. It is involved in protein, nucleic acid, carbohydrate, and lipid metabolism, as well as in the control of gene transcription, growth, development, and differentiation. SLC39A6 belongs to a subfamily of proteins that show structural characteristics of zinc transporters.SLC39A6 (Solute Carrier Family 39 Member 6) is a Protein Coding gene. Diseases associated with SLC39A6 include Acrodermatitis Enteropathica and Breast Cancer. Among its related pathways are Metal ion SLC transporters and Transport of inorganic cations/anions and amino acids/oligopeptides. Gene Ontology (GO) annotations related to this gene include metal ion transmembrane transporter activity and zinc ion transmembrane transporter activity.

Specifications

Catalog Number	KC-0875
Cell Line Name	293T-SLC39A6-KO-3B3-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone with human SLC39A6 gene knockout, No.3B3
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T-SLC39A6-KO-3B3 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of 293T-SLC39A6-KO-3B3 cell line stable clone using PCR sequencing.

Figure 2: Characterization of 293T-SLC39A6-KO-3B3 cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:3-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Taylor KM, Nicholson RI. The LZT proteins; the LIV-1 subfamily of zinc transporters. Biochim Biophys Acta. 2003 Apr 1;1611(1-2):16-30. doi: 10.1016/s0005-2736(03)00048-8. PMID: 12659941.
Zhu X, Yu C, Wu W, Shi L, Jiang C, Wang L, Ding Z, Liu Y. Zinc transporter ZIP12 maintains zinc homeostasis and protects spermatogonia from oxidative stress during spermatogenesis. Reprod Biol Endocrinol. 2022 Jan 22;20(1):17. doi: 10.1186/s12958-022-00893-7. PMID: 35065654; PMCID: PMC8783530.
Fertal SA, Poterala JE, Ponik SM, Wisinski KB. Stromal Characteristics and Impact on New Therapies for Metastatic Triple-Negative Breast Cancer. Cancers (Basel). 2022 Feb 27;14(5):1238. doi: 10.3390/cancers14051238. PMID: 35267548; PMCID: PMC8909697.