- Synonyms:
- 293[HEK-293] dCas9-KRAB
- Description:
- Parent cell line: 293 (also known as HEK-293)
- Stably expresses dCas9-KRAB (created by knocking-in a KRAB-dCas9 (from Streptococcus pyogenes) expression cassette into AAVS1 safe harbor locus using CRISPR/Cas9 gene editing technology)
- This cell line also expresses red fluorescence protein (RFP) and hygromycin resistance gene.
- Keywords:
- Culture Type: Cell line
- Origin/Source:
- Organism:
- Species: Human (Homo sapiens)
- Age: Fetus
- Tissue of Origin: Embryonic Kidney
- Organism:
- Cell line features:
- Morphology: Epithelial-like
- Cell Type: Epithelial
- Growth Mode: Adherent
- Morphology: Epithelial-like
- Applications:
- Can be used to carry out CRISPRi-RNA guided gene expression repression (knock-down)
- Can be used to carry out CRISPRi-RNA guided gene expression repression (knock-down)
- Handling and maintenance:
- Biosafety level: BSL 2 (It may differ in your country, follow the guidelines of your institute/country)
- Complete culture medium:
- EMEM + 10% FBS + 100 µg/mL hygromycin
- EMEM + 10% FBS + 100 µg/mL hygromycin
- Culture conditions:
- Temperature: Humidified 37°C incubator
- Atmosphere: 5% CO₂, 95% Air
- Subculturing:
- Cell dissociation procedure: 0.25% (w/v) Trypsin- 0.53 mM EDTA solution
- Harvesting time: 80% confluence culture
- Subcultivation Ratio: 1:6 to 1:10
- Medium Renewal: 2 to 3 times per week
- Seeding densities: 1 x 104 – 4 x 104 viable cells/cm2
- Cell dissociation procedure: 0.25% (w/v) Trypsin- 0.53 mM EDTA solution
- Cryopreservation
- Freezing medium: EMEM + 10% (v/v) FBS + 5% (v/v) DMSO
- Cryopreservation temperature: Liquid nitrogen gas phase or -150°C freezer
- Availability:
- American Type Culture Collection (ATCC): Product link
- References:
- https://www.atcc.org/products/crl-1573dcas9-krab
- Brown et al., 2014. Seven Strains of Enterovirus D68 Detected in the United States during the 2014 Severe Respiratory Disease Outbreak. Genome Announc 2(6): e01201-14, 2014. PMID-25414503; Full-Text Links: ASM, PMC
- Abbreviations
- EMEM (Eagle’s Modified Eagle’s Medium)
- FBS (Fetal Bovine Serum)
- DMSO (Dimethyl sulfoxide)
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