GNPAT KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 1 bp insertion in exon 3.
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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, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 1 bp insertion in exon 3
Alternative names
Acyl CoA dihydroxyacetonephosphateacyltransferase, Acyl-CoA:dihydroxyacetonephosphateacyltransferase, DAP-AT, DHAP-AT, Dihydroxyacetone phosphate acyltransferase, EC 2.3.1.42, GNPAT_HUMAN, Glycerone-phosphate O-acyltransferase, OTTHUMP00000036147
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GNPAT KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 1 bp insertion in exon 3.
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, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 1 bp insertion in exon 3
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
GNPAT
- 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
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255928). 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.
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
GNPAT also known as glyceronephosphate O-acyltransferase or dihydroxyacetone phosphate acyltransferase (DAPAT) is an enzyme that plays an important role in lipid metabolism. The enzyme has a mass of approximately 73 kDa. It is expressed mainly in peroxisomes organelles involved in various metabolic functions. The enzyme catalyzes the initial and rate-limiting step in the biosynthesis of ether lipids like plasmalogens by converting dihydroxyacetone phosphate (DHAP) to acyl-DHAP.
- Biological function summary
GNPAT is essential in lipid biosynthesis since ether lipids are integral components of cell membranes and signaling molecules. The enzyme does not work in isolation; it forms part of a multi-enzyme complex that includes other important proteins like alkylglycerone phosphate synthase (AGPS). This association allows GNPAT to efficiently catalyze the first steps in the formation of plasmalogens which serve key roles in protecting cells from oxidative stress and in cell signaling.
- Pathways
GNPAT functions efficiently within the ether lipid synthesis pathway which is critical for normal cellular membrane composition and function. The activity of GNPAT impacts the levels of plasmalogens influencing pathways involved in membrane dynamics and antioxidant defense. It works closely with AGPS enhancing the production of essential ether lipids and affects phospholipid metabolism interfacing indirectly with proteins involved in peroxisomal and mitochondrial functions.
- Associated diseases and disorders
GNPAT dysfunction has associations with rhizomelic chondrodysplasia punctata (RCDP) a genetic disorder characterized by skeletal abnormalities and developmental delay. The enzyme's interaction with proteins like Phospholipase A2 is relevant in understanding its role in RCDP. Additionally alterations in plasmalogen levels due to impaired GNPAT function are observed in neurological conditions such as Alzheimer's disease. This connection highlights the importance of GNPAT in maintaining normal brain lipid metabolism and preventing cognitive decline.
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4 product images
Sanger Sequencing - Human GNPAT (DAPAT) knockout HeLa cell line (ab265611)
Allele-1: 10 bp deletion in exon 3.
Sanger Sequencing - Human GNPAT (DAPAT) knockout HeLa cell line (ab265611)
Allele-2: 1 bp deletion in exon 3.
Sanger Sequencing - Human GNPAT (DAPAT) knockout HeLa cell line (ab265611)
Allele-3: 1 bp insertion in exon 3.
Sanger Sequencing - Human GNPAT (DAPAT) knockout HeLa cell line (ab265611)
Sequencing chromatogram displaying sequence edit in exon 3
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