HNRNPF KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 4 and 13 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 4 and 13 bp deletion in exon 4 and Insertion of the selection cassette in exon 4
Recommended products
HNRNPF KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 4 and 13 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 4 and 13 bp deletion in exon 4 and Insertion of the selection cassette in exon 4
- Concentration
Properties
- Gene name
- HNRNPF
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
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 short-term storage conditions
- -196°C
- 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 HEK293T cell line (ab255449). 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
Heterogeneous nuclear ribonucleoprotein F (hnRNP F) also known as hnRNP-F or HNRNPF plays an important role in RNA processing within the nucleus. It belongs to the family of hnRNPs which bind to nascent pre-mRNA molecules influencing their stability and transport. HnRNP F has a molecular weight of approximately 46 kDa. It is ubiquitously expressed found in various tissues but it exhibits higher levels of expression in the testis and brain. Its structured domains allow it to bind to RNA with high specificity impacting several RNA metabolic processes directly.
- Biological function summary
HnRNP F interacts with RNA molecules to modulate alternative splicing mRNA stability and transport. It often forms part of larger ribonucleoprotein complexes that assemble on pre-mRNA. These complexes facilitate the accurate processing of pre-mRNA by guiding the necessary splicing machinery. HnRNP F commonly associates with other proteins such as hnRNP H enhancing its ability to regulate splice site selection and influencing mRNA diversity and function within the cell.
- Pathways
HnRNP F is importantly involved in the regulation of gene expression through the mRNA splicing pathway. It particularly affects the spliceosome assembly a multi-protein complex essential for the removal of introns. hnRNP F's involvement in this pathway intersects with proteins such as SF2/ASF and U2AF65 which are key regulators in splicing. Furthermore hnRNP F influences the response to cellular stress pathways by stabilizing specific mRNAs under stress conditions often cooperating with proteins involved in stress granules.
- Associated diseases and disorders
HnRNP F has significant associations with neurodegenerative diseases and cancer. In neurodegenerative diseases aberrant splicing events influenced by hnRNP F are linked to conditions such as Alzheimer's disease where it interacts with splicing factors like SRSF1. In cancer altered expression or malfunction of hnRNP F contributes to splicing variants that promote oncogenesis notably in association with proteins like MYC which drives cancer cell proliferation. These connections make hnRNP F a potential target for therapeutic interventions in disease treatment.
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3 product images
Sanger Sequencing - Human HNRNPF knockout HEK-293T cell line (ab266333)
Allele-1: Insertion of the selection cassette in exon 4
Sanger Sequencing - Human HNRNPF knockout HEK-293T cell line (ab266333)
Allele-2: 13 bp deletion in exon 4.
Sanger Sequencing - Human HNRNPF knockout HEK-293T cell line (ab266333)
Allele-3: 11 bp deletion in exon 4.
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