KC-4357

COLO205-XRN2-KO-5C4(++/-)-Cell-Line

44741

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Background of COLO205-XRN2-KO-5C4(++/-)-Cell-Line

XRN2 is a 5'-to-3' exoribonuclease that is predominantly localized in the nucleus. By degrading or trimming various classes of RNA, XRN2 contributes to essential processes in gene expression such as transcription termination and ribosome biogenesis. Despite limited substrate specificity in vitro, XRN2 targets a specific subset of RNA by interacting with other proteins in cells.

Specifications

Catalog Number	KC-4357
Cell Line Name	COLO205-XRN2-KO-5C4(++/-)-Cell-Line
Host Cell Line	COLO205
Description	Stable COLO205 clone with human XRN2 gene knockout, No.5C4
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+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 26.76 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

COLO205-XRN2-KO-5C4 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of COLO205-XRN2-KO-5C4 cell line stable clone using PCR sequencing.

Figure 2: Characterization of COLO205-XRN2-KO-5C4 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of COLO205-XRN2-KO-5C4 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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Miki TS, Großhans H. The multifunctional RNase XRN2. Biochem Soc Trans. 2013 Aug;41(4):825-30. doi: 10.1042/BST20130001. PMID: 23863139.
2.Aygün I, Miki TS. Nuclear RNA Regulation by XRN2 and XTBD Family Proteins. Cell Struct Funct. 2021 Nov 6;46(2):73-78. doi: 10.1247/csf.21041. Epub 2021 Sep 3. PMID: 34483148; PMCID: PMC10511037.