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AB277816

Human GLI2 knockout A549 cell line

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GLI2 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

CJS, GLI Kruppel family member GLI2, GLI family zinc finger 2, GLI2_HUMAN, Glioma associated oncogene family zinc finger, HPE9, Oncogene GLI2, PHS 2, THP, THP1, THP2, Tax helper protein, Tax helper protein 1, Tax helper protein 2, Tax responsive element 2 holding protein, Tax responsive element 25 bp sequence binding protein, Zinc finger protein GLI2

1 Images
Next Generation Sequencing - Human GLI2 knockout A549 cell line (AB277816)
  • NGS

Lab

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

142 bp deletion after Ser402

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Disease

Carcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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 (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.

Properties and storage information

Gene name
GLI2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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