KC-6100

293T-SLITRK6-Cell-Line

60704

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Background of 293T-SLITRK6-Cell-Line

SLITRK6 encodes a member of the SLITRK protein family. Members of this family are integral membrane proteins that are characterized by two N-terminal leucine-rich repeat (LRR) domains and a C-terminal region that shares homology with trk neurotrophin receptors. This protein functions as a regulator of neurite outgrowth required for normal hearing and vision. Mutations in this gene are a cause of myopia and deafness.

Specifications

Catalog Number	KC-6100
Cell Line Name	293T-SLITRK6-Cell-Line
NCBI/UniProt Accession Number	NM_032229.2
Clone Number	7#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous SLITRK6 gene
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	70% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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

Cell Line Generation

293T-SLITRK6-cell-line was generated using a lentiviral vector expressing the SLITRK6 sequence.

Characterization

Figure 1: Characterization of SLITRK6 overexpression in the 293T-SLITRK6 stable clone using qPCR.

Figure 2: Characterization of SLITRK6 in the 293T -SLITRK6 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

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

References

1.Yu F, Zhao X, Li M, Meng M. SLITRK6 promotes the progression of lung adenocarcinoma by regulating PI3K/AKT/mTOR signaling and Warburg effect. Apoptosis. 2023 Aug;28(7-8):1216-1225. doi: 10.1007/s10495-023-01838-0. Epub 2023 May 23. PMID: 37219677.
2.Liu X, Liu Y, Liu Z, Zhang Y, Ma Y, Bai J, Yao H, Wang Y, Zhao X, Li R, Song X, Chen Y, Feng Z, Wang L. Identification of SLITRK6 as a Novel Biomarker in hepatocellular carcinoma by comprehensive bioinformatic analysis. Biochem Biophys Rep. 2021 Oct 27;28:101157. doi: 10.1016/j.bbrep.2021.101157. PMID: 34754951; PMCID: PMC8564567.
3.Morlet T, Rabinowitz MR, Looney LR, Riegner T, Greenwood LA, Sherman EA, Achilly N, Zhu A, Yoo E, O'Reilly RC, Jinks RN, Puffenberger EG, Heaps A, Morton H, Strauss KA. A homozygous SLITRK6 nonsense mutation is associated with progressive auditory neuropathy in humans. Laryngoscope. 2014 Mar;124(3):E95-103. doi: 10.1002/lary.24361. Epub 2013 Dec 17. PMID: 23946138; PMCID: PMC3925201.