KC-6600

JJN-3-VAV3-HiBiT-KI(+/-) Cell Line

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Background of JJN-3-VAV3-HiBiT-KI(+/-) Cell Line

The VAV3 gene, located on chromosome 1p13.3, encodes a guanine nucleotide exchange factor (GEF) of the VAV family. VAV3 functions by catalyzing GDP-GTP exchange on Rho GTPases (e.g., RAC1, RHOA), thereby regulating cytoskeletal reorganization, proliferation, and migration. Its expression spans hematopoietic and non-hematopoietic tissues, underscoring its systemic role in signaling transduction. Dysregulation of VAV3 is implicated in several pathological conditions. For instance, in castration-resistant prostate cancer (CRPC), VAV3 potentiates androgen receptor (AR) activity, driving therapy resistance. Moreover, VAV3 overexpression in breast cancer, glioblastoma, and leukemia promotes epithelial-mesenchymal transition (EMT), metastasis, and chemoresistance. Consequently, VAV3 has emerged as a therapeutic target, with strategies including small-molecule inhibitors and RNA-based approaches to overcome resistance in malignancies.

Specifications

Catalog Number	KC-6600
Cell Line Name	JJN-3-VAV3-HiBiT-KI(+/-) Cell Line
NCBI/UniProt Accession Number	10451/Q9UKW4
Clone Number	1A4
Host Cell Line	JJN-3
Description	JJN-3 cell line stable clone with endogenous HiBiT-tagged VAV3 gene
Quantity	One vial 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/IMDM(1:1) + 20% FBS
Selection Marker	NA
Morphology	Lymphoblast
Subculture	Split saturated culture 1:3-1:4 every 3-4 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

JJN-3-VAV3-HiBiT-KI(+/-) cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of JJN-3-VAV3-HiBiT-KI(+/-) cell line stable clone using PCR sequencing.

Figure 2: Characterization of JJN-3-VAV3-HiBiT-KI(+/-) cell line stable clone using RT-PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM/IMDM(1:1) + 20% 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 1:3-1:4 every 3-4 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

Al-Hawary SIS, Najm MA, Hjazi A, et al. VAV3 in human cancers: Mechanism and clinical implication. Pathol Res Pract. 2023 Aug;248:154681. doi: 10.1016/j.prp.2023.154681. Epub 2023 Jul 13. PMID: 37467637.
Wang Y, Li Y, Zhang X, et al. Unravelling the roles of Vav3 in cytoskeletal control and angiogenesis. Life Sci. 2025;360:123347. doi: 10.1016/j.lfs.2024.123347. Epub 2024 Dec 20. PMID: 40618919.

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.