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AB266214

Human USP16 knockout HEK-293T cell line

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USP16 KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 2 and 5 bp deletion in exon 2. 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

Deubiquitinating enzyme 16, Human ubiquitin processing protease, MSTP039, UBP M, UBP16_HUMAN, Ubiquitin Specific Protease 16, Ubiquitin Specific peptidase 16, Ubiquitin carboxyl-terminal hydrolase 16, Ubiquitin thiolesterase 16, Ubiquitin-processing protease UBP-M, Ubiquitin-specific-processing protease 16

3 Images
Sanger Sequencing - Human USP16 knockout HEK-293T cell line (AB266214)
  • Sanger seq

Unknown

Sanger Sequencing - Human USP16 knockout HEK-293T cell line (AB266214)

Allele-1 : 5 bp deletion in exon2

Sanger Sequencing - Human USP16 knockout HEK-293T cell line (AB266214)
  • Sanger seq

Lab

Sanger Sequencing - Human USP16 knockout HEK-293T cell line (AB266214)

Sequencing chromatogram displaying sequence edit in exon 2

Sanger Sequencing - Human USP16 knockout HEK-293T cell line (AB266214)
  • Sanger seq

Unknown

Sanger Sequencing - Human USP16 knockout HEK-293T cell line (AB266214)

Allele-2 : 4 bp deletion in exon 2.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 2 and 5 bp deletion in exon 2

Product details

Recommended control: Human wild-type HEK293T 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.

Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Culture medium: DMEM (High Glucose) + 10% FBS

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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
  • A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
  • Cells should be passaged when they have achieved 80-90% confluence.

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
USP16
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Handling procedures

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.

USP16 also known as Ubiquitin Specific Peptidase 16 functions as a deubiquitinating enzyme that regulates protein degradation by cleaving ubiquitin from ubiquitin-conjugated protein substrates. It has a molecular mass of approximately 97 kDa. USP16 is expressed in various tissues with high levels in the brain and heart. It has an integral role in maintaining protein homeostasis by regulating degradation pathways necessary for cellular function.
Biological function summary

The enzymatic activity of USP16 removes ubiquitin moieties contributing to the stabilization of target proteins. It participates in several cellular processes including cell cycle regulation DNA damage response and chromatin remodeling. USP16 is associated with the Polycomb group (PcG) complex which plays a role in transcriptional repression. This involvement in epigenetic regulation highlights its importance in gene expression and cellular differentiation.

Pathways

USP16 is significant in the Wnt signaling pathway and the Hedgehog signaling pathway. In the Wnt pathway USP16 regulates beta-catenin degradation influencing cell proliferation and differentiation. Through the Hedgehog pathway USP16 impacts the stability of GLI transcription factors affecting developmental processes and tissue patterning. USP16 interacts with proteins like Axin and APC in the Wnt pathway illustrating its role in complex regulatory networks.

Researchers have found connections between USP16 dysregulation and cancer particularly leukemia and neurodegenerative diseases like Alzheimer's disease. In cancer USP16 overexpression can lead to increased cell proliferation through pathways involving proteins such as MYC and Cyclin D1. Related to neurodegenerative disorders USP16's interaction with proteins like amyloid precursor protein (APP) contributes to pathologies like Alzheimer's where proteostasis and ubiquitination play critical roles. Understanding USP16's functions and connections can provide insights into therapeutic targets for these diseases.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Viability

~ 80%

Product protocols

Product promise

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