KC-3703

MDA-MB-231-STING-KO-2A2-Cell-Line

Stable MDA-MB-231 clone with human STING gene knockout, No.2A2

40702

Home » MDA-MB-231-STING-KO-2A2-Cell-Line

Background of MDA-MB-231-STING-KO-2A2-Cell-Line

STING encodes a five transmembrane protein that functions as a major regulator of the innate immune response to viral and bacterial infections. It is the facilitator of innate immune signaling that acts as a sensor of cytosolic DNA from bacteria and viruses and promotes low production of type I interferon. STING acts by binding cyclic dinucleotides: recognizes and binds cyclic di-GMP (c-di-GMP), a second messenger produced by bacteria, and cyclic GMP-AMP (cGAMP), a messenger produced by cGAS in response to DNA virus in the cytosol.

Specifications

Catalog Number	KC-3703
Cell Line Name	MDA-MB-231-STING-KO-2A2-Cell-Line
Host Cell Line	MDA-MB-231
Description	Stable MDA-MB-231 clone with human STING gene knockout, No.2A2
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 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 38 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

MDA-MB-231-STING-KO-2A2 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of MDA-MB-231-STING-KO-2A2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of MDA-MB-231-STING-KO-2A2 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of MDA-MB-231-STING-KO-2A2 cell line stable clone using western blot.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 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 1:3-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 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

Samson N, Ablasser A. The cGAS-STING pathway and cancer. Nat Cancer. 2022 Dec;3(12):1452-1463. doi: 10.1038/s43018-022-00468-w. Epub 2022 Dec 12. PMID: 36510011.
Xie J, Li Y, Shen X, Goh G, Zhu Y, Cui J, Wang LF, Shi ZL, Zhou P. Dampened STING-Dependent Interferon Activation in Bats. Cell Host Microbe. 2018 Mar 14;23(3):297-301.e4. doi: 10.1016/j.chom.2018.01.006. Epub 2018 Feb 22. PMID: 29478775; PMCID: PMC7104992.
Zhang R, Qin X, Yang Y, Zhu X, Zhao S, Zhang Z, Su Q, Zhao Z, Yin X, Meng X, Zhang Z, Li Y. STING1 is essential for an RNA-virus triggered autophagy. Autophagy. 2022 Apr;18(4):816-828. doi: 10.1080/15548627.2021.1959086. Epub 2021 Aug 2. PMID: 34338134; PMCID: PMC9037512.

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.