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AB289221

Human FOXP3 knockout HCT116 cell line

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FOXP3 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

View Alternative Names

AIID, DIETER, FOXP3_HUMAN, FOXP3delta7, Forkhead box P3, Forkhead box protein P3, IPEX, Immune dysregulation polyendocrinopathy enteropathy X linked, Immunodeficiency polyendocrinopathy enteropathy X linked, JM2, MGC141961, MGC141963, OTTHUMP00000025832, OTTHUMP00000025833, OTTHUMP00000226737, PIDX, Scurfin, XPID

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Next Generation Sequencing - Human FOXP3 knockout HCT116 cell line (AB289221)
  • NGS

Lab

Next Generation Sequencing - Human FOXP3 knockout HCT116 cell line (AB289221)

44 bp deletion after Ala 110 of WT protein

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

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 HCT116 cell line (ab288559). 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.
There is evidence in literature (PMID: 28591725) that detects the expression of FOXP3 in WT HCT116 cell line.

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
FOXP3
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.
Culture medium

McCoY5a + 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.

FOXP3 also recognized as Forkhead box P3 is a transcription factor critical for immune system function. It weighs approximately 47 kDa. FOXP3 is mainly expressed in regulatory T cells (Tregs) which play a role in maintaining immune tolerance. Scientists primarily identify its presence in the nucleus where it facilitates transcriptional regulation. Methods like 'FOXP3 IHC' and 'FOXP3 IHC mouse' are useful for detecting this protein in tissue samples while 'FOXP3 antibodies' aid in research on this target. Techniques like 'FOXP3 staining' and 'FOXP3 flow cytometry' further allow for the analysis of its expression and function.
Biological function summary

This transcription factor operates by regulating the expression of a variety of genes involved in Treg cell development and function. FOXP3 can interact with other proteins to form complexes that modulate gene expression. Its role is essential in enabling Tregs to suppress immune responses a process necessary for preventing autoimmunity. The interaction extends to other key regulators within Tregs such as the protein complex 221D which may have overlapping functional responsibilities. 'FOXP3 236a/E7' reflects its genetic variants detectable in different studies providing insights into its biological functions.

Pathways

FOXP3 is a significant player within the immune regulation pathway. It integrates into the TGF-beta signaling pathway in which it controls the expression of genes important for Treg functions. The protein recruits other transcription factors like NF-kB coordinating intricate network regulations within immune cells. FOXP3's activity orchestrates the balance between immune activation and suppression ensuring a well-functioning immune response that prevents overreactions leading to autoimmune conditions.

FOXP3 mutations or dysregulation can link to autoimmune diseases like immune dysregulation polyendocrinopathy enteropathy X-linked syndrome (IPEX). This transcription factor's malfunction implicates cancer where inadequate Treg function can either suppress antitumor immunity or result in tumor progression. Proteins such as CTLA-4 which is also involved in immune regulation connect to FOXP3 in these contexts emphasizing FOXP3’s significant role in maintaining immune homeostasis and its impact on disease development.

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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