KC-6057

MDA-MB-231-luc2-DHPS-KO(+/-) Cell Line

65462

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Background of MDA-MB-231-luc2-DHPS-KO(+/-) Cell Line

Deoxyhypusine synthase (DHPS) catalyzes the rate-limiting hypusination of eukaryotic translation initiation factor 5A (eIF5A), a unique modification essential for eIF5A function. Recent DHPS knockout studies have uncovered its diverse roles: in ovarian cancer, CRISPR/Cas9 knockout suppresses tumor growth and metastasis by blocking EMT and TGFβ signaling; in neurodevelopment, neuron-specific Dhps knockout in mice causes more severe growth, neurodevelopmental, and cognitive deficits than Eif5a knockout, while zebrafish dhps knockdown recapitulates microcephaly and epileptiform activity of DHPS deficiency syndrome; in pancreatic development, DHPS loss impairs β‑cell mass and pancreatic growth, with eIF5A knockout showing minimal effects, suggesting DHPS may act independently of eIF5A hypusination. Thus, DHPS knockout serves as a powerful tool for studying hypusination biology and DHPS‑related diseases.

Specifications

Catalog Number	KC-6057
Cell Line Name	MDA-MB-231-luc2-DHPS-KO(+/-) Cell Line
Clone Number	2A4
Host Cell Line	MDA-MB-231-luc2
Description	Stable MDA-MB-231-luc2 cell line with human DHPS gene knockout
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	RPMI1640 + 10%FBS + 400μg/mL Hygromycin B + 10% DMSO
Propagation Medium	RPMI1640 + 10%FBS + 400μg/mL Hygromycin B
Selection Marker	NA
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:2-1:4 every 2-3 days; seed out at about 1-3 x 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 38 hours
Mycoplasma Status	Negative

Cell Line Generation

MDA-MB-231-luc2-DHPS-KO(+/-) cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of MDA-MB-231-luc2-DHPS-KO(+/-) Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of MDA-MB-231-luc2-DHPS-KO(+/-) Cell Line stable clone using RT-PCR sequencing.

Figure 3: Characterization of MDA-MB-231-luc2-DHPS-KO(+/-) cell line stable clone using western blot.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10%FBS + 400μg/mL Hygromycin B)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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (RPMI1640 + 10%FBS + 400μg/mL Hygromycin B + 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

Zhao G, Zhao X, Liu Z, Wang B, Dong P, Watari H, Pfeffer LM, Tigyi G, Zhang W, Yue J. Knockout or inhibition of DHPS suppresses ovarian tumor growth and metastasis by attenuating the TGFβ pathway. Sci Rep. 2025 Jan 6;15(1):917. doi: 10.1038/s41598-025-85466-5. PMID: 39762448; PMCID: PMC11704301.
Anderson CM, Kulkarni A, Maier B, Huang F, Figatner K, Chakraborty A, Pratuangtham S, May SC, Tersey SA, Anderson RM, Mirmira RG. Hypusinated and unhypusinated isoforms of the translation factor eIF5A exert distinct effects in models of pancreas development and function. J Biol Chem. 2025 Feb;301(2):108209. doi: 10.1016/j.jbc.2025.108209. Epub 2025 Jan 18. PMID: 39832654; PMCID: PMC11869520.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.