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AB266702

Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line

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RNASEH2A 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 1 and 1 bp insertion in exon 1.

View Alternative Names

AGS4, Aicardi-Goutieres syndrome 4 protein, EC 3.1.26.4, RNASEH2A, RNASEHI, RNH2A_HUMAN, RNHIA, RNHL, RNase H(35), RNase H2 subunit A, RNase HI large subunit, Ribonuclease H2 subunit A, Ribonuclease HI large subunit, Ribonuclease HI subunit A

3 Images
Sanger Sequencing - Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line (AB266702)
  • Sanger seq

Unknown

Sanger Sequencing - Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line (AB266702)

Allele-1 : 1 bp deletion in exon 1

Sanger Sequencing - Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line (AB266702)
  • Sanger seq

Unknown

Sanger Sequencing - Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line (AB266702)

Allele-2 : 1 bp insertion in exon 1.

Sanger Sequencing - Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line (AB266702)
  • Sanger seq

Lab

Sanger Sequencing - Human RNASEH2A (Ribonuclease H2, subunit A) knockout HEK-293T cell line (AB266702)

Sequencing chromatogram displaying sequence edit in exon 1

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, 1 bp deletion in exon 1 and 1 bp insertion in exon 1

Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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, 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
RNASEH2A
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.

Ribonuclease H2 subunit A also known as RNASEH2A is one part of the RNase H2 enzyme complex. This protein weighs approximately 33 kilodaltons and plays an important role in RNA-DNA hybrid processing. RNASEH2A operates by cleaving the RNA strand of RNA-DNA hybrids which is necessary for maintaining genome stability. The protein expresses in a variety of tissues. Importantly it is found in high levels in cells with rapid division like embryonic and stem cells.
Biological function summary

RNASEH2A facilitates the removal of ribonucleotides from DNA which helps in DNA replication and repair. As a part of the RNase H2 complex alongside subunits B and C it ensures the correct removal of embedded ribonucleotides from DNA strands. This process is important for maintaining the integrity of the genome and preventing mutations. The activity of RNASEH2A directly impacts DNA replication fidelity and overall genome stability.

Pathways

RNASEH2A plays an important role in the maintenance of genome integrity and the DNA repair pathway. It directly interacts with the process of ribonucleotide excision repair which prevents genomic instability. RNASEH2A works alongside enzymes involved in DNA replication and repair mechanisms like DNA polymerases to correct DNA strands. The collaboration of these proteins ensures the accuracy of genomic information passed during cell division.

RNASEH2A is associated with Aicardi-Goutières syndrome a severe neurodevelopmental disorder. This syndrome results from mutations in any of the RNase H2 subunits including RNASEH2A leading to accumulated DNA damage and a chronic interferon response. Another condition linked to malfunctions in this protein is systemic lupus erythematosus where an abnormal immune response attacks the body's own cells. Both conditions connect RNASEH2A with the regulation of immune responses and highlight its role in preventing inappropriate autoimmunity.

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

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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