GNC5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. KO cell line generated by CRISPR/Cas9 system. Individual gRNAs were subcloned into pSpCas9 (BB)-2A-Puro and then stably transfected into U2OS cells. Transfected cells were selected by puromycin.
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Publications
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- Genes & development 33:1751-17742019Systematic bromodomain protein screens identify homologous recombination and R-loop suppression pathways involved in genome integrity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJae Jin Kim et. al.PubMed 31753913
Key facts
- Cell type
- U-2 OS
- Species or organism
- Human
- Tissue
- Bone
- Form
- Liquid
- Knockout validation
- Western blot
- Mutation description
- KO cell line generated by CRISPR/Cas9 system. Individual gRNAs were subcloned into pSpCas9 (BB)-2A-Puro and then stably transfected into U2OS cells. Transfected cells were selected by puromycin.
Alternative names
1110051E14Rik, AW212720, EC 2.3.1.48, GCN5 (general control of amino-acid synthesis, yeast, homolog)-like 2, GCN5 general control of amino-acid synthesis 5-like 2 (yeast), GCN5L2, General control of amino acid synthesis protein 5-like 2, General control of amino acid synthesis, yeast, homolog-like 2, HGCN5, Histone acetyltransferase GCN5, Histone acetyltransferase KAT2A, K(lysine) acetyltransferase 2A, KAT2A_HUMAN, Lysine acetyltransferase 2A, MGC102791, PCAF-b, STAF97, general control of amino acid synthesis 5-like 2, hsGCN5
Recommended products
GNC5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. KO cell line generated by CRISPR/Cas9 system. Individual gRNAs were subcloned into pSpCas9 (BB)-2A-Puro and then stably transfected into U2OS cells. Transfected cells were selected by puromycin.
Key facts
- Cell type
- U-2 OS
- Species or organism
- Human
- Tissue
- Bone
- Form
- Liquid
- Knockout validation
- Western blot
- Mutation description
- KO cell line generated by CRISPR/Cas9 system. Individual gRNAs were subcloned into pSpCas9 (BB)-2A-Puro and then stably transfected into U2OS cells. Transfected cells were selected by puromycin.
- Disease
- Osteosarcoma
- Concentration
Properties
- Gene name
- GNC5
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Western blot
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 + 10% FBS + 1% Penicillin-Streptomycin-Glutamine
- 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 duration
- A few days
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
For more information about this cell line please see publication: PMID: 31753913.
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 U2-OS cell line (ab300225). 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 + 10% FBS + 1% Penicillin-Streptomycin-Glutamine
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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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
KAT2A also known as GCN5 is a lysine acetyltransferase enzyme with a molecular mass of approximately 92 kDa. It mainly acetylates histone proteins and affects transcriptional regulation by modifying chromatin structure. KAT2A/GCN5 is expressed in various tissues with significant expression in the nucleus of eukaryotic cells. Its role as a histone acetyltransferase places it as an important player in transcriptional activation by modifying the chromatin to a more open conformation facilitating access by transcription machinery.
- Biological function summary
KAT2A/GCN5 participates in several cellular processes as part of larger protein complexes. It is a component of the SAGA (Spt-Ada-Gcn5 acetyltransferase) and TFTC (TBP-free TAF-containing complex) both involved in transcription regulation. Through these complexes KAT2A influences gene expression by acetylating histone residues which in turn affects the recruitment of other transcriptional activators or repressors. This activity also links it to non-histone substrates that support DNA damage repair and cellular senescence.
- Pathways
KAT2A/GCN5 is integrally connected to the regulation of transcription and chromatin remodeling pathways. It interacts closely with the transcription factor c-Myc assisting in cell cycle regulation and proliferation. Additionally its presence in the p53 signaling pathway highlights its involvement in controlling cell growth and apoptosis. KAT2A/GCN5 activity influences downstream events in these pathways by affecting the acetylation state of specific histones and non-histone proteins.
- Associated diseases and disorders
The dysregulation of KAT2A/GCN5 is linked to cancer particularly in breast and colorectal cancers. Its role in acetylating key proteins like c-Myc and affecting pathways like p53 can lead to unchecked cell growth when improperly regulated. Furthermore KAT2A is connected to neurodegenerative diseases through its interactions with other histone modifiers such as HDAC1 and HDAC2 implicating it in conditions where neural cell function or maintenance goes awry.
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1 product image
Western blot - Human GNC5 knockout U2-OS cell line (ab300217)
Levels of endogenous protein were analyzed by western blot. Please see publication for more detail: PMID: 31753913.
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