KC-4253

22Rv1 CHD7 KO 1C2 Cell Line

43427

Home » 22Rv1 CHD7 KO 1C2 Cell Line

Background of 22Rv1 CHD7 KO 1C2 Cell Line

The CHD7 gene, encoding the chromodomain helicase DNA-binding protein 7, is a multifunctional protein that plays a critical role in chromatin remodeling, transcriptional regulation, and signal transduction. CHD7 is a member of the CHD family of proteins, characterized by the presence of chromodomains and a SNF2-related helicase/ATPase domain. This protein is involved in a variety of developmental processes and is essential for neural crest development and sensory organ formation.Mutations in the CHD7 gene are primarily associated with CHARGE syndrome, a complex multisystem disorder characterized by a constellation of features including coloboma of the eye, heart defects, choanal atresia, retarded growth and development, genital hypoplasia, and ear anomalies. CHARGE syndrome exhibits a high degree of phenotypic variability, even among individuals with the same CHD7 mutation.

Specifications

Catalog Number	KC-4253
Cell Line Name	22Rv1 CHD7 KO 1C2 Cell Line
Host Cell Line	22Rv1
Description	Stable 22Rv1 clone with human CHD7 gene knockout, No.1C2
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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

22Rv1-CHD7-KO-1C2 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of 22Rv1-CHD7-KO-1C2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of 22Rv1-CHD7-KO-1C2 cell line stable clone using RT-PCR sequencing.

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

Vissers, L. J., et al. (2004). CHARGE syndrome caused by mutations in CHD7.Nature Genetics, 36(12), 1210-1212. https://doi.org/10.1038/ng1464
Mehaffey, C. A., et al. (2015). CHD7 regulates neural crest development through direct control of the BMP pathway.Nature Communications, 6, 7559. https://doi.org/10.1038/ncomms8559
Shankar, S., et al. (2017). CHD7 regulates multiple signaling pathways during inner ear hair cell development.Developmental Biology, 424(1), 33-44. https://doi.org/10.1016/j.ydbio.2017.01.018

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.