CDA KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1
Recommended products
CDA KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion 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, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
CDA
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Cell culture
- Biosafety level
EU: 2 US: 2
- Viability
~ 80%
- 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 long-term storage conditions
-196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
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
Cytidine deaminase (CDA) also known as CDD is a zinc-dependent enzyme responsible for deaminating cytidine. The full form of CDA has a molecular mass of approximately 41 kDa. CDA is widely expressed in various tissues including liver kidney and leukocytes. It plays an important role in nucleotide metabolism by converting cytidine to uridine and 2'-deoxycytidine to 2'-deoxyuridine which impacts nucleic acid stability and synthesis. Researchers frequently use techniques such as ELISA and Western blotting to study CDA expression and activity often referencing it in contexts like "CDA Western" or "57 seconds CDA."
- Biological function summary
CDA enzyme influences the pyrimidine salvage pathway by facilitating nucleotide degradation and recycling. It operates independently rather than as part of a protein complex. The activity of this enzyme ensures a balanced nucleotide pool essential for DNA replication and repair processes. This balance affects cellular proliferation and survival highlighting the importance of studying CDA protein in normal physiology and various pathological conditions.
- Pathways
CDA contributes significantly to pyrimidine metabolism and its recycling pathway. It maintains a connection with ribonucleotide reductase which plays a role in the regulation of deoxyribonucleotide pools during DNA synthesis. This interplay highlights the enzyme’s involvement in DNA repair pathways ensuring proper cell division and genomic integrity. Within these pathways enzymes like cytidine triphosphate synthetase also relate to CDA by participating in nucleotide homeostasis.
- Associated diseases and disorders
Improper CDA function is linked to specific cancers including acute myeloid leukemia due to altered nucleotide metabolism leading to genomic instability. The enzyme's activity also affects the pharmacokinetics of cytidine analogues like gemcitabine used in cancer treatment. CDA overexpression might result in reduced drug efficacy thereby hindering therapy in affected patients. Connections exist between CDA and proteins involved in chemoresistance such as ribonucleotide reductase showing its significance in treatment outcomes.
Product promise
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Sanger Sequencing - Human CDA knockout HeLa cell line (ab264848)
Allele-1: 1 bp insertion in exon 1.
Sanger Sequencing - Human CDA knockout HeLa cell line (ab264848)
Allele-2: Insertion of the selection cassette in exon 1.
Sanger Sequencing - Human CDA knockout HeLa cell line (ab264848)
Allele-3: Insertion of the selection cassette in exon 1.
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