RFWD3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2.
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Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2
Alternative names
BC027246, E3 ubiquitin-protein ligase RFWD3, FLJ10520, MGC27888, RFWD3_HUMAN, RING finger and WD repeat domain-containing protein 3, RING finger protein 201, RNF201, Ring finger and WD repeat domain 3
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RFWD3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- RFWD3
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
RFWD3 also known as RNF201 is an E3 ubiquitin-protein ligase with a mass of approximately 75 kDa. This protein adds ubiquitin to specific substrate proteins marking them for degradation by the proteasome. RFWD3 is mainly expressed in the cell nucleus where it plays a role in maintaining genomic stability. The ubiquitination process mediated by RFWD3 is critical for its function in DNA repair mechanisms especially in response to DNA damage.
- Biological function summary
RFWD3 is key in the context of DNA repair and cell cycle regulation. It acts in conjunction with other repair proteins forming complexes necessary for homologous recombination repair. This protein interacts with replication protein A (RPA) during DNA damage response facilitating repair by driving proper assembly and disassembly of DNA repair complexes. RFWD3 ensures the preservation of genomic fidelity and cell viability under conditions of stress.
- Pathways
RFWD3 plays a central role in DNA damage response and repair mechanisms. It is directly involved in the homologous recombination pathway working alongside proteins such as CHK1 and BRCA1. RFWD3 contributes to the stability of these pathways enabling error-free repair of DNA double-strand breaks. The protein's function in these pathways is critical for the prevention of chromosomal aberrations and cancerous transformations.
- Associated diseases and disorders
RFWD3 has been linked to cancer and Fanconi anemia. Defects in RFWD3 can lead to impaired DNA repair processes resulting in genomic instability and promoting carcinogenesis. Additionally it plays a role in the Fanconi anemia pathway interacting with proteins like FANCD2 and FANCI. This interaction is essential for proper DNA crosslink repair and deficiencies can contribute to the development of Fanconi anemia characterized by bone marrow failure and increased cancer risk.
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1 product image
Sanger Sequencing - Human RFWD3 knockout HeLa cell line (ab265128)
Homozygous: 2 bp deletion in exon 2.
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