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AB264746

Human SIAH1 knockout HeLa cell line

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SIAH1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 4 bp deletion in exon 2. 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 SIAH1 knockout HeLa cell line (AB264746)
  • Sanger seq

Unknown

Sanger Sequencing - Human SIAH1 knockout HeLa cell line (AB264746)

Allele-2 : 2 bp deletion in exon 2.

Sanger Sequencing - Human SIAH1 knockout HeLa cell line (AB264746)
  • Sanger seq

Unknown

Sanger Sequencing - Human SIAH1 knockout HeLa cell line (AB264746)

Allele-1 : 4 bp deletion in exon 2.

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 2 and 4 bp deletion in exon 2

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
SIAH1
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.

SIAH1 also known as Seven In Absentia Homolog 1 functions mechanically as a RING-type E3 ubiquitin-protein ligase. This protein weighs approximately 32 kDa and adds ubiquitin to target proteins tagging them for proteasomal degradation. Researchers have found SIAH1 expressed in various tissue types including brain heart and kidney. By regulating protein stability SIAH1 plays a significant role in maintaining cellular protein homeostasis.
Biological function summary

SIAH1 influences cell cycle control and apoptosis by interacting with components of protein complexes. It associates with the ubiquitin-proteasome pathway to mediate degradation of key regulatory proteins. SIAH1 helps in reducing cellular proliferation and enhances apoptotic signaling which are vital processes for maintaining balance between cell survival and death. As part of these processes studies have connected SIAH1 to modulating the fate of proteins involved in cell cycle arrest.

Pathways

Several interactions of SIAH1 contribute to signal transduction and apoptosis. One significant pathway involving SIAH1 is the p53 signaling pathway. SIAH1 regulates the degradation of proteins within this pathway including p21 and cyclin D1 which influence cellular responses to DNA damage. Additionally SIAH1 interacts with β-catenin linking it to the Wnt signaling pathway. This interaction affects the regulation of cell growth and differentiation.

SIAH1 has links to cancer and neurodegenerative diseases. In cancer altered SIAH1 expression affects tumor suppressor activity through its interaction with p53 which can lead to uncontrolled cell growth. Moreover SIAH1 connects to neurodegenerative disorders like Parkinson's disease where abnormal protein degradation contributes to neuronal death. In such contexts researchers observe altered interactions between SIAH1 and proteins like parkin a ubiquitin ligase also implicated in 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

Product protocols

Product promise

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