KC-4762

293T-ACVR1C-KO Cell Line

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Background of 293T-ACVR1C-KO Cell Line

The Activin A Receptor Type 1C (ACVR1C), also known as ALK-7, belongs to the transforming growth factor-beta (TGF-β) superfamily of serine/threonine kinase receptors. ACVR1C plays a significant role in various biological processes, including cell proliferation, differentiation, and apoptosis by mediating signals from activin A, a cytokine involved in numerous physiological functions. Research has indicated that ACVR1C is implicated in metabolic regulation, immune responses, and embryonic development. Dysregulation or mutations in this gene have been linked to several diseases, such as polycystic ovary syndrome (PCOS) and cancer. Studies on ACVR1C have revealed its potential as a therapeutic target for these conditions due to its influence on intracellular signaling pathways like SMAD-dependent and -independent pathways. Understanding the molecular mechanisms underlying ACVR1C's function could provide new insights into disease pathogenesis and pave the way for innovative treatments.

Specifications

Catalog Number	KC-4762
Cell Line Name	293T-ACVR1C-KO Cell Line
Host Cell Line	293T
Description	Stable 293T clone with ACVR1C gene knockout, No.2C2
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
Selection Marker	NA
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-ACVR1C-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of ACVR1C knockout in 293T using PCR sequencing.

Figure 2: Characterization of ACVR1C knockout in 293T using RT-PCR sequencing.

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 1:4-1:8 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

Ibáñez CF. Regulation of metabolic homeostasis by the TGF-β superfamily receptor ALK7. FEBS J. 2022 Oct;289(19):5776-5797. doi: 10.1111/febs.16090. Epub 2021 Jul 11. PMID: 34173336.
Lin SY, Huang H, Yu JJ, Su F, Jiang T, Zhang SY, Lv L, Long T, Pan HW, Qi JQ, Zhou Q, Tang WF, Ding GW, Wang LM, Tan LJ, Yin J. Activin A receptor type 1C single nucleotide polymorphisms associated with esophageal squamous cell carcinoma risk in Chinese population. World J Gastrointest Oncol. 2025 Jan 15;17(1):96702. doi: 10.4251/wjgo.v17.i1.96702. PMID: 39817119; PMCID: PMC11664604.
Hong X, Wen B, Zhang H, Li Y, Wu H, Zhao W, Luo X. Biological effects of NODAL on endometrial cancer cells and its underlying mechanisms. Exp Ther Med. 2021 Apr;21(4):402. doi: 10.3892/etm.2021.9833. Epub 2021 Feb 25. PMID: 33717261; PMCID: PMC7938447.
https://www.ncbi.nlm.nih.gov/gene/130399