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AB265402

Human SPG11 knockout HeLa cell line

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SPG11 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion 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 SPG11 knockout HeLa cell line (AB265402)
  • Sanger seq

Unknown

Sanger Sequencing - Human SPG11 knockout HeLa cell line (AB265402)

Allele-1 : 2 bp deletion in exon 1.

Sanger Sequencing - Human SPG11 knockout HeLa cell line (AB265402)
  • Sanger seq

Unknown

Sanger Sequencing - Human SPG11 knockout HeLa cell line (AB265402)

Allele-2 : 1 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, 1 bp deletion in exon 1 and 2 bp deletion 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 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.

What's included?

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Properties and storage information

Gene name
SPG11
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.

SPG11 also known as spatacsin is a protein with a mass of approximately 249 kDa. This protein is expressed mainly in neural tissues including the brain and spinal cord. Mechanically SPG11 interacts with various other proteins contributing to the maintenance and function of the lysosomal compartment within cells. It plays an important role in membrane trafficking processes particularly in the vesicles involved in endocytosis and autophagy.
Biological function summary

In the context of neuronal health SPG11 is significant for the function of neurons. It forms part of a larger protein complex known as the AP-5 adaptin complex which assists in lysosomal positioning and function. By ensuring proper lysosomal function SPG11 supports neuronal maintenance and health. Issues with its function disrupt lysosomal degradation pathways and can have broad implications for neuron viability.

Pathways

SPG11 is essential in endolysosomal and autophagic pathways serving as a connection point for effective lysosomal transport. These pathways include interactions with proteins such as AP5Z1 and ZFYVE26 which collaborate to modulate lysosomal positioning and maintenance. SPG11's proper operation within these pathways allows for efficient cellular digestion of macromolecules and removal of damaged organelles critical for cellular health.

Mutations in SPG11 lead to rare neurological conditions like spastic paraplegia type 11 and some forms of juvenile amyotrophic lateral sclerosis (ALS). SPG11's mutation or dysfunction can result in improper lysosomal degradation contributing to the nervous system's degeneration. Related proteins like spastizin (ZFYVE26) and AP5Z1 when disrupted also exhibit similar symptoms of neural decay highlighting SPG11's integral role in neuronal disease 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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