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AB265139

Human GRK4 knockout HeLa cell line

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GRK4 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 9 and 31 bp deletion in exon 9.

View Alternative Names

G protein-coupled receptor kinase 2 (Drosophila)-like, G protein-coupled receptor kinase 4, G protein-coupled receptor kinase GRK4, G proteinm coupled receptor kinase GRK4, GPRK2L, GPRK4, GRK4_HUMAN, GRK4a, ITI1

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Sanger Sequencing - Human GRK4 knockout HeLa cell line (AB265139)
  • Sanger seq

Unknown

Sanger Sequencing - Human GRK4 knockout HeLa cell line (AB265139)

Allele-1 : 31 bp deletion in exon 9.

Sanger Sequencing - Human GRK4 knockout HeLa cell line (AB265139)
  • Sanger seq

Unknown

Sanger Sequencing - Human GRK4 knockout HeLa cell line (AB265139)

Allele-2 : 1 bp deletion in exon 9.

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 9 and 31 bp deletion in exon 9

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 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
GRK4
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.

The GRK4 or G protein-coupled receptor kinase 4 belongs to the family of serine/threonine kinases. It plays a mechanical role in phosphorylating activated G protein-coupled receptors (GPCRs) which leads to receptor desensitization. GRK4 has a molecular mass of about 66 kDa and is widely expressed in tissues including the kidney testis and brain. Such expression pattern suggests its involvement in numerous physiological processes.
Biological function summary

G protein-coupled receptor kinase 4 regulates GPCR signaling by facilitating receptor internalization and degradation. This action modulates cellular responsiveness to hormones and neurotransmitters. GRK4 does not form part of a complex but its regulatory function directly impacts GPCR pathways. GRK4 achieves this control by adding phosphate groups typically functioning as a feedback mechanism during receptor activity.

Pathways

GRK4 serves as an important regulator in the MAPK signaling pathway and the renin-angiotensin-aldosterone system (RAAS). In MAPK pathways GRK4 helps mediate signal transduction by controlling receptor activity. It also interacts with other GRKs like GRK2 to fine-tune cellular responses and maintain homeostasis. Its involvement in RAAS highlights its significance in cardiovascular regulation.

G protein-coupled receptor kinase 4 has strong links to hypertension and Parkinson's disease. Its dysregulation or genetic variants can lead to abnormal blood pressure regulation by altering dopamine receptors via RAAS. In Parkinson's disease GRK4's influence on dopamine-related pathways suggests a connection to the condition's neurological symptoms. Aberrations in GRK4 activities may also impact interactions with proteins like GRK6 contributing to disease progression.

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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