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ADAR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 2 and 1 bp insertion in exon 2.

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Images

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846), expandable thumbnail
  • Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846), expandable thumbnail
  • Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846), expandable thumbnail
  • Sanger Sequencing - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Human ADAR (ADAR1) knockout HEK-293T cell line (AB266846), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 2 and 1 bp insertion in exon 2

Alternative names

136 kDa double-stranded RNA-binding protein, ADAR, AGS6, AV242451, Adenosine deaminase RNA specific, Adenosine deaminase RNA specific 1, Adenosine deaminase acting on RNA 1 A, Adenosine deaminase that act on RNA, DRADA, DSRAD_HUMAN, Double-stranded RNA-specific adenosine deaminase, Double-stranded RNA-specific editase Adar, Dsh, EC 3.5.4.-, G1P1, Ifi4 protein, Interferon induced protein 4, Interferon-inducible protein 4, K88DSRBP, P136, Pre-mRNA adenosine deaminase, RNA adenosine deaminase 1, RNA-editing deaminase 1, RNA-editing enzyme 1, dsRNA adenosine deaminase, mZaADAR

Recommended products

ADAR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 2 and 1 bp insertion in exon 2.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 2 and 1 bp insertion in exon 2
Concentration
Loading...

Properties

Gene name
ADAR
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: 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 HEK293T cell line (Human wild-type HEK-293T 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.

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

ADAR1 also known as RNA-specific adenosine deaminase 1 or ADAR is an enzyme with a mass of approximately 150 kDa. This protein targets double-stranded RNA (dsRNA) and acts mechanistically to convert adenosine to inosine in pre-mRNA sequences a process known as A-to-I RNA editing. ADAR1 is expressed in numerous tissues with high levels in the brain liver and lungs. Its localization within cells can vary often found in both the nucleus and cytoplasm which influences its function.

Biological function summary

ADAR1 plays a role in the regulation of RNA molecules affecting their stability and translation. It is linked to the editosome complex working alongside other proteins to perform RNA editing tasks. By modifying the coding potential of mRNAs ADAR1 contributes to the diversity of proteomes and helps manage the responses to viral RNAs giving the immune system tools to recognize endogenous and exogenous RNA.

Pathways

ADAR1 is significant in the interferon signaling pathway and RNA processing pathways. It operates in coordination with proteins like PKR which is involved in the response to viral infections. ADAR1 ensures that the immune response is not directed against the self highlighting its role in the regulation of the innate immune system. These pathways are critical in maintaining homeostasis and preventing unchecked immune responses.

Associated diseases and disorders

Mutations or dysregulation of ADAR1 have associations with autoimmune diseases like Aicardi-Goutieres syndrome and certain cancers. The enzyme's role in editing RNA makes it essential in preventing inappropriate immune attacks against the body's own cells highlighting its interaction with MDA5 another protein involved in immune regulation. Understanding ADAR1 and its related pathways may offer potential therapeutic targets for these conditions including exploration into ADAR1 inhibitors as interventions.

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Product protocols

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

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