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AB264947

Human POLE4 knockout HeLa cell line

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POLE4 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 1. 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 POLE4 knockout HeLa cell line (AB264947)
  • Sanger seq

Unknown

Sanger Sequencing - Human POLE4 knockout HeLa cell line (AB264947)

Homozygous : Insertion of the selection cassette 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, Homozygous: Insertion of the selection cassette in exon 1

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
POLE4
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 biological target POLE4 also known as DNA polymerase epsilon subunit 4 is an important component of the DNA replication machinery. This protein has a molecular weight of approximately 10 kDa. POLE4 primarily participates in the function of DNA polymerase epsilon—an important enzyme necessary for synthesizing the leading strand during DNA replication. The expression of POLE4 usually happens in the nucleus where DNA replication takes place indicating its essential role in this cellular location.
Biological function summary

DNA polymerase epsilon subunit 4 supports both DNA replication and repair mechanisms. POLE4 is a part of the larger DNA polymerase epsilon complex which includes other subunits such as POLE POLE2 and POLE3. In concert this complex ensures high-fidelity DNA synthesis and checks for errors during replication. This process is critical for maintaining genome stability and preventing mutations that could lead to severe cellular outcomes.

Pathways

The function of DNA polymerase epsilon involving POLE4 places it within the DNA replication and cell cycle pathways. In these pathways POLE4 works closely with POLE and other associated elements to facilitate the proper progression of the replication fork and the overall cell cycle. The association with these pathways highlights the protein's involvement in ensuring that cells divide accurately and efficiently with minimal errors.

Abnormal function or expression of POLE4 can connect to conditions such as cancer and genomic instability disorders. These diseases may involve POLE as well given its direct interactive role within the same complex. Therefore studying POLE4 in the context of these conditions could reveal insights into how errors in DNA replication and repair can contribute to the development and progression of such 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

Product protocols

Product promise

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