KC-1261

Ba/F3-KRAS-G12V Cell Line

63919

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Background of Ba/F3-KRAS-G12V Cell Line

KRAS (Kirsten rat sarcoma viral oncogene homolog), also known as KRAS2, RASK2, or NS3, is a small GTPase encoded by the KRAS gene on chromosome 12p12.1. As a member of the RAS superfamily, KRAS functions as a molecular switch cycling between active GTP-bound and inactive GDP-bound states to regulate cell proliferation, differentiation, and survival. KRAS is ubiquitously expressed but exhibits high mutation frequency in non-small cell lung cancer (NSCLC), colorectal cancer (CRC), and pancreatic ductal adenocarcinoma (PDAC). The G12V mutation (glycine to valine at codon 12) impairs intrinsic GTP hydrolysis, locking KRAS in a constitutively active state, and accounts for approximately 29% of PDAC and 10% of CRC cases. While G12C-targeted inhibitors like sotorasib and adagrasib are FDA-approved, G12V has historically been considered undruggable. However, RMC-5127, a RAS(ON) G12V-selective inhibitor, entered Phase 1 clinical trial in 2026 for advanced solid tumors harboring G12V, representing a major advance for this common mutation.

Specifications

Catalog Number	KC-1261
Cell Line Name	Ba/F3-KRAS-G12V Cell Line
NCBI/UniProt Accession Number	NM_033360
Host Cell Line	Ba/F3 Cell Line
Description	Stable Ba/F3 cell line expressing exogenous human KRAS harboring the G12V mutation.
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% basal medium+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS
Selection Marker	Puromycin
Morphology	Suspension, round
Subculture	Split saturated culture 1:10 every 3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Ba/F3-KRAS-G12V cell line was generated using a retroviral vector expressing the human KRAS sequence harboring the G12V mutation.

Characterization

Figure 1: Characterization of KRAS-G12V in the Ba/F3-KRAS-G12V stable clone using PCR sequencing.

Figure 2: Assessment of the in vitro inhibitory effects of AMG510 and MRTX849 on the proliferation of Ba/F3-KRAS-G12V cells using a cell proliferation assay.

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:10 every 3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Wang, Yunpeng et al. “Case Report: Anlotinib combined with PD-1 inhibitor and sequential GA regimen or FOLFIRINOX Chemotherapy in treatment of KRAS G12V mutated pancreatic ductal adenocarcinoma with liver metastasis: A case and literature review.” Frontiers in immunology vol. 13 1016647. 13 Oct. 2022, doi:10.3389/fimmu.2022.1016647
2. Zhou, Xile et al. “Multiple Strategies to Develop Small Molecular KRAS Directly Bound Inhibitors.” Molecules (Basel, Switzerland) vol. 28,8 3615. 21 Apr. 2023, doi:10.3390/molecules28083615
3. Rathod, Lala S et al. “KRAS inhibitors in drug resistance and potential for combination therapy.” Tumori vol. 111,1 (2025): 20-40. doi:10.1177/03008916241289206

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.