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AB266211

Human RFX5 knockout HEK-293T cell line

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RFX5 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion 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'.

View Alternative Names

DNA-binding protein RFX5, Influences HLA class II expression, RFX5_HUMAN, Regulatory factor X 5, Regulatory factor X subunit 5, Regulatory factor X, 5 (influences HLA class II expression), Rfx5 protein

1 Images
Sanger Sequencing - Human RFX5 knockout HEK-293T cell line (AB266211)
  • Sanger seq

Unknown

Sanger Sequencing - Human RFX5 knockout HEK-293T cell line (AB266211)

Homozygous : 1 bp insertion in exon 3

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: 1 bp insertion 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
RFX5
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.

RFX5 also known as Regulatory Factor X 5 is a transcription factor characterized by a mass of approximately 74 kDa. It plays an essential role in the modulation and transcriptional regulation of genes. RFX5 is predominantly expressed in the nucleus of human cells where it binds to DNA and regulates gene expression. This protein is vital to the regulation of major histocompatibility complex (MHC) class II genes affecting how cells present antigens.
Biological function summary

RFX5 functions as part of the RFX complex an important element for the transcription of MHC class II genes. It interacts with other regulatory factor X proteins to form this complex enabling the activation of target gene transcription. RFX5 acts by binding to the X box motifs within the promoters of the MHC class II genes. Its presence is necessary to maintain the immune response capability by ensuring the expression of these genes which is a fundamental process in antigen presentation to immune cells.

Pathways

RFX5 integrates into the MHC class II antigen presentation pathway. It has a significant relationship with the CIITA transcriptional regulator with CIITA enhancing the activity of RFX5 during the assembly of the MHC class II transcriptional machinery. Another important pathway is the immune response pathway where RFX5 is important in coordinating the expression of genes that help the immune system recognize and respond to pathogens effectively.

Deficiencies or malfunctions in RFX5 have strong connections with Bare Lymphocyte Syndrome (BLS) type II marked by reduced expression of MHC class II molecules. In this context RFX5's interactions with CIITA are critical because improper function of either can lead to compromised immune responses. Studies also suggest an association between disrupted RFX5 activity and autoimmune diseases where improper antigen presentation can lead to the immune system attacking the body's own tissues.

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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