NTCC? HEK293/dCas9-BFP-KRAB Cell Line

dCas9基因編輯穩(wěn)定表達(dá)細(xì)胞株

Catalog number: NTCC?-SL371

Product: HEK293 cell line stably expressing CRISPR dCas9 and KRAB

Description: This product is a cell line stably expressing the catalytically inactive, HA-taggeddCas9 nuclease fused to blue fluorescent protein (BFP) and the transcriptionalrepression domain Krüppel associated box (KRAB). This cell line also containsblasticidin resistance gene. The target guide RNAs can be transfected ortransduced in the cell line to repress the gene transcription.Quantity: 1 vial of 2 x 106cells; frozenShipping conditions: Dry iceStorage conditions: Liquid nitrogen vapor phase. Remove the item from the dry ice packaging andcheck all items for damage and leakage. Place immediately into storage at orbelow -140°C, preferably into the liquid nitrogen vapor phase, until use.Source of parental line:HEK293Organism: Homo sapiens, humanTissue: embryonic kidneyCell type: epithelialSafety instructions: To ensure safety, protective gloves, clothing, and a face mask should be wornwhen handling frozen vials. Some leakage of liquid nitrogen into the vial mayoccur during storage. The liquid nitrogen will be converted to gas upon thawing.Due to the nature of nitrogen gas, pressure may build within the vial and possiblyresult in the vial exploding or losing its cap. This may cause flying debris.Thawing procedure: The vial of cells should be thawed in a 37°C water bath with gentle agitation. Foroptimal performance, the vial should be thawed in under two minutes. Ensurethat the cap of the vial did not loosen upon thawing, and re-tighten as needed.Spray the vial with 70% EtOH and wipe off. Repeat. Using aseptic technique, addthe contents of the vial to 9ml of complete growth medium (without selection).Centrifuge for 5min at 125xg.Aspirate the medium, being careful not to disturbthe pellet. Re-suspend in 10mL of complete growth medium, and place into aculture vessel of your choice. Only add selection to the medium after 24 hours inculture.

Culture conditions:

Cryopreservation:

Quality control:

Complete Growth Medium：The base medium for this cell line is DMEM. For optimal growth and maintenanceof selection, add the following components to the base medium: fetal bovineserum to a final concentration of 10%.SelectionBlasticidin to a final concentration of 20 ug/mLCulture temperature:37°C with 5% CO2Subculture:Rinse the cells with PBS without cations, digest cells with 0.25% (w/v) TrypsinEDTA (0.53mM) solution and split at 1:6 to 1:10 ratio.Freeze slowly in complete growth medium supplemented with 5% (v/v) DMSO.The genome integration, expression levels of dCas9-BFP-KRAB, and thetranscription repression of target genes using the cell line have been verified byqPCR. All other QC specifications have also been met. For detailed information,please refer to the certificate of analysis.