RNF149 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
Images
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, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1
Alternative names
DNA polymerase-transactivated protein 2, DNAPTP2, E3 ubiquitin-protein ligase RNF149, RING finger protein 149, RN149_HUMAN, Rnf149
Recommended products
RNF149 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
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, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- RNF149
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
RNF149 also known as RING Finger Protein 149 is an E3 ubiquitin-protein ligase which means it catalyzes the transfer of ubiquitin from an E2 ubiquitin-conjugating enzyme to a substrate protein tagging it for degradation. The mass of RNF149 is approximately 49 kDa. It can be found expressed in various tissues with notable presence in the liver and kidney. This protein contains a RING-type zinc finger domain which is critical for its ligase activity and substrate specificity.
- Biological function summary
RNF149 participates in protein quality control by mediating the ubiquitination of proteins that require degradation therefore maintaining protein homeostasis. It acts independently and is not part of a larger protein complex. By tagging damaged or misfolded proteins for degradation RNF149 contributes to the cellular stress response ensuring that cells respond to potential stressors effectively and efficiently.
- Pathways
The involvement of RNF149 in the ubiquitin-proteasome system places it within the protein degradation pathway which is essential for regulating protein turnover and cellular function. It interacts with several other proteins including ubiquitin-conjugating enzyme E2s in this pathway. RNF149 may also intersect with apoptosis pathways due to its role in controlling the degradation of apoptotic regulators linking it to proteins involved in cell death processes.
- Associated diseases and disorders
RNF149 shows a connection to certain cancers and neurodegenerative diseases. Abnormal expression or mutations in RNF149 can relate to the deregulation of protein turnover contributing to the development of hepatocellular carcinoma. It is also connected to neurodegenerative disorders due to its role in degrading damaged proteins with connections to proteins like p53 in cancer pathways and tau proteins in neurological contexts.
Product promise
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2 product images
Sanger Sequencing - Human RNF149 knockout HeLa cell line (ab265671)
Allele-2: Insertion of the selection cassette in exon 1.
Sanger Sequencing - Human RNF149 knockout HeLa cell line (ab265671)
Allele-1: 1 bp deletion in exon 1.
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