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AB266049

Human TTC3 knockout HeLa cell line

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TTC3 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Sanger Sequencing - Human TTC3 knockout HeLa cell line (AB266049)
  • Sanger seq

Unknown

Sanger Sequencing - Human TTC3 knockout HeLa cell line (AB266049)

Homozygous : Insertion of the selection cassette in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2

Disease

Adenocarcinoma

Product details

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.

What's included?

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Properties and storage information

Gene name
TTC3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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

DMEM (High Glucose) + 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.

The TTC3 protein also known as Tetratricopeptide Repeat Domain 3 has a molecular weight of approximately 215 kDa. This protein functions as an E3 ubiquitin-protein ligase which means it plays a role in tagging proteins for degradation via the ubiquitin-proteasome system. TTC3 is widely expressed in the brain especially within neurons and is also found in other tissues such as the heart and kidneys. It contains tetratricopeptide repeats which are known for facilitating protein-protein interactions essential for its role in cellular function and signaling.
Biological function summary

TTC3 interacts in various processes associated with cell development and stability. It is not part of a larger protein complex but operates notably in cell cycle progression and neuronal differentiation. TTC3 regulates the stability of proteins by adding ubiquitin molecules which are small regulatory proteins to target proteins. This activity is important for maintaining the cellular environment especially in tissues where precise control over protein levels is necessary.

Pathways

Many cellular pathways are influenced by TTC3 including the PI3K-AKT signaling pathway which mediates several cellular processes such as growth and survival. TTC3 has connections with proteins such as AKT1 through this pathway which helps regulate cell proliferation and neurogenesis. Additionally TTC3 is involved in the stability regulation of cyclin proteins influencing the cell cycle by interacting with components of the cyclin-dependent kinase (CDK) pathway therefore affecting cell cycle checkpoints and transitions.

Evidence shows that changes in TTC3 expression have links to neurological conditions like Down syndrome and Alzheimer's disease. In these disorders altered TTC3 function affects neuronal growth and function likely through dysregulated ubiquitination processes. Additionally TTC3 interacts with proteins like DYRK1A which is also implicated in neurodegenerative conditions and Down syndrome pathophysiology. Altered TTC3 activity therefore contributes to the molecular mechanisms driving these diseases suggesting its potential as a therapeutic target.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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