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AB265725

Human UNG knockout HeLa cell line

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UNG KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

View Alternative Names

DGU, DKFZp781L1143, HIGM 4, OTTHUMP00000240527, OTTHUMP00000240528, OTTHUMP00000240529, UDG, UNG 1, UNG 15, UNG2, UNG_HUMAN, Uracil DNA glycosylase 1, Uracil DNA glycosylase 2, Uracil-DNA glycosylase

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Sanger Sequencing - Human UNG knockout HeLa cell line (AB265725)
  • Sanger seq

Unknown

Sanger Sequencing - Human UNG knockout HeLa cell line (AB265725)

Allele-2 : Insertion of the selection cassette in exon 1.

Sanger Sequencing - Human UNG knockout HeLa cell line (AB265725)
  • Sanger seq

Unknown

Sanger Sequencing - Human UNG knockout HeLa cell line (AB265725)

Allele-1 : 2 bp deletion in exon 1.

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, 2 bp deletion in exon 1 and Insertion of the selection cassette in exon 1

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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, 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
UNG
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

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.

UNG which stands for Uracil-DNA glycosylase is an enzyme that functions mechanically to remove uracil from DNA. This preventive action is necessary because uracil can result from the deamination of cytosine or due to misincorporation during DNA replication. UNG exhibits a mass of approximately 33 kDa and shows expression in various tissues underlining its importance across different cellular environments. UNG is ubiquitously expressed but displays higher activity in proliferating cells making it integral during cell division.
Biological function summary

URACIL-DNA glycosylase engages in base excision repair (BER) an important DNA repair process. This enzyme initiates the repair by excising uracil bases creating an abasic site subsequently processed by other repair proteins. UNG actively functions within single-enzymatic action but also associates with various DNA repair complexes highlighting its multifaceted role in genome maintenance. It guards against mutations by maintaining DNA integrity reducing the risk errors pose to genetic stability.

Pathways

UNG plays a role in DNA repair and cell cycle pathways contributing essential actions to genomic stability and cell proliferation. Specifically it integrates into the base excision repair pathway working alongside other proteins like APEX1 and DNA polymerase beta to ensure proper repair of damaged DNA. These related proteins cooperate to fix abasic sites that arise after UNG's glycosylase activity ensuring that genetic information remains accurate and undamaged as cells replicate.

Mutations in UNG can present increased susceptibility to immunodeficiency and cancer. One such disorder includes hyper-IgM syndrome a condition where BER defects lead to an impaired immune response. Proteins like CD40 and CD40L intersect with UNG-related pathways impacting germinal center reactions and immunoglobulin isotype switching. The enzyme's dysfunction potentially contributes to the accumulation of genetic errors fostering oncogenesis in several cancers by allowing abnormal cells to escape normal regulatory mechanisms.

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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