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GLI2 KO cell line available now. KO validated by Next Generation Sequencing. Free of charge wild type control available.

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Next Generation Sequencing - Human GLI2 knockout A549 cell line (AB277816), expandable thumbnail

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

Knockout validation

Next Generation Sequencing

Alternative names

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GLI2 KO cell line available now. KO validated by Next Generation Sequencing. Free of charge wild type control available.

Alternative names

Key facts

Cell type

A549

Form

Liquid

Disease

Carcinoma

Concentration
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Properties

Gene name

GLI2

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing

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 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 A549 cell line (Human wild-type A549 cell line ab288558). 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 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

Activity summary

Gli2 also known as GLI family zinc finger 2 is a transcription factor with a molecular mass of approximately 160 kDa. It is an important component of the Hedgehog (Hh) signaling pathway. Gli2 primarily resides in the cytoplasm and migrates to the nucleus upon activation. Expression of Gli2 is widespread but most notable in developing tissues and adult brain. In the nucleus it regulates transcription by binding to specific DNA sequences.

Biological function summary

Gli2 acts as an activator or repressor of gene expression depending on the cellular context. It forms part of the Gli family of transcription factors playing a significant role in cellular growth differentiation and tissue patterning. While not usually part of stable protein complexes Gli2 interacts transiently with other proteins such as Suppressor of Fused (SuFu) to regulate its transcriptional activity. This interaction influences the processing and degradation of Gli proteins.

Pathways

The Hedgehog signaling cascade prominently features Gli2 as a critical mediator. This pathway is fundamental in embryonic development. Alongside other Gli proteins like Gli1 and Gli3 Gli2 helps to translate extracellular Hh signals into precise cellular responses. In the absence of Hh ligands Gli2 remains tethered in the cytoplasm in a complex with SuFu limiting its transcription activity. Upon Hh pathway activation Gli2 translocates to the nucleus and modulates target gene expression.

Associated diseases and disorders

Gli2 has a notable connection to cancers including basal cell carcinoma and medulloblastoma. Overactive Hedgehog signaling often involves dysregulation of Gli2 which promotes tumorigenesis by driving inappropriate cellular proliferation. Additionally Gli2 links to the genetic disorder Greig cephalopolysyndactyly syndrome via its interactions with Gli3 protein. These connections underline the importance of Gli2 as a therapeutic target in certain pathological conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

1 product image

  • Next Generation Sequencing - Human GLI2 knockout A549 cell line (ab277816), expandable thumbnail

    Next Generation Sequencing - Human GLI2 knockout A549 cell line (ab277816)

    142 bp deletion after Ser402

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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'.

For licensing inquiries, please contact partnerships@abcam.com