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TREX1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 4 bp insertion in exon 1.

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Images

Western blot - Human TREX1 knockout A549 cell line (AB266927), expandable thumbnail
  • Cell Culture - Human TREX1 knockout A549 cell line (AB266927), expandable thumbnail
  • Sanger Sequencing - Human TREX1 knockout A549 cell line (AB266927), expandable thumbnail
  • Sanger Sequencing - Human TREX1 knockout A549 cell line (AB266927), expandable thumbnail

Key facts

Cell type
A549
Species or organism
Human
Tissue
Lung
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 4 bp insertion in exon 1

Alternative names

Recommended products

TREX1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 4 bp insertion in exon 1.

Key facts

Cell type
A549
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 4 bp insertion in exon 1
Disease
Carcinoma
Concentration
Loading...

Properties

Gene name
TREX1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male

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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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 A549 cell line (ab255450). 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

TREX1 also known as Three Prime Repair Exonuclease 1 acts as a DNA exonuclease which degrades single-stranded DNA from the 3' end. This protein exhibits a mass of 33 kDa. TREX1's expression is largely found in the cytoplasm of many cell types including those in the immune system and various tissues like the liver spleen and heart. Through its enzymatic activity TREX1 helps maintain genomic stability by processing DNA intermediates that arise from incomplete DNA replication or repair processes.

Biological function summary

TREX1 removes excessive DNA that might trigger immune responses. It is not a part of a large protein complex but works closely with substrates involved in DNA repair and replication. By degrading abnormal DNA TREX1 prevents the initiation of inappropriate immune responses that could result in autoimmunity. Its functionality is essential in preventing the cell from converting these DNA fragments into ligands for DNA sensors which could activate immune signaling pathways.

Pathways

TREX1 plays a significant role in the DNA damage response and innate immune pathways. In the DNA damage response pathway TREX1 functions alongside proteins like ATR and ATM which address DNA replication stress and repair. Within the innate immune pathway it interacts with cGAS (cyclic GMP-AMP synthase) which can become activated in response to cytosolic DNA leading to the production of type I interferons - powerful immune modulators.

Associated diseases and disorders

Mutations in TREX1 have been linked to autoimmune diseases such as Aicardi-Goutières syndrome and systemic lupus erythematosus. In these conditions the altered or deficient TREX1 leads to the accumulation of DNA in the cytoplasm falsely triggering an antiviral immune response. This relationship with cGAS is critical as the cGAS-STING pathway becomes activated due to the presence of unprocessed DNA resulting in chronic inflammation and contributing to the autoimmune pathology observed in these disorders.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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