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AB266150

Human PEX5 (PER3) knockout HEK-293T cell line

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PEX5 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 3. 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 PEX5 (PER3) knockout HEK-293T cell line (AB266150)
  • Sanger seq

Unknown

Sanger Sequencing - Human PEX5 (PER3) knockout HEK-293T cell line (AB266150)

Homozygous : Insertion of the selection cassette in exon3

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, Homozygous: Insertion of the selection cassette in exon 3

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
PEX5
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.

PEX5 (Peroxisomal Biogenesis Factor 5) also referred to as peroxin-5 functions as a cytosolic receptor for peroxisomal targeting signal 1 (PTS1) proteins. With a mass of approximately 71 kDa PEX5 exhibits widespread expression in various tissues. It facilitates the transport of proteins into peroxisomes which are cellular organelles critical for lipid metabolism and the detoxification of reactive oxygen species. PEX5 achieves this by binding to PTS1-tagged proteins in the cytosol and guiding them to the peroxisomal membrane.
Biological function summary

The PEX5 protein plays a significant role in peroxisomal protein import by participating in a transient multi-protein transport complex at the peroxisome membrane. This complex ensures that imported proteins are released into the peroxisomal matrix. PEX5 interacts closely with PEX14 a peroxisomal membrane protein to facilitate the docking of PTS1-carrying proteins. PEX5 is indispensable for maintaining normal peroxisomal function and integrity as it ensures that specific enzymes reach their destination to carry out metabolic processes.

Pathways

PEX5 interacts within the broader lipid metabolism and reactive oxygen species detoxification pathways. It contributes to the proper functioning of enzymes involved in beta-oxidation of fatty acids and plasmalogen synthesis. PEX5 operates in conjunction with the PEX7 receptor which handles the PTS2 pathway helping to manage the trafficking of enzymes important for peroxisomal operations thereby integrating lipid metabolism with cellular homeostasis.

Mutations in the PEX5 gene often result in peroxisome biogenesis disorders such as Zellweger syndrome which is part of Zellweger spectrum disorders. These conditions manifest due to defects in peroxisomal function leading to neurological impairment and metabolic dysregulation. PEX14 has been identified as a related protein as it cooperates with PEX5 in the peroxisomal matrix protein import pathway. Understanding PEX5's function and interactions is essential for developing strategies to address these genetic disorders.

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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