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AB265454

Human QTRT1 knockout HeLa cell line

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QTRT1 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and 46 bp deletion in exon 1.

View Alternative Names

FP3235, Guanine insertion enzyme, Queuine tRNA ribosyltransferase 1, Queuine tRNA-ribosyltransferase, TGT, 43 KD subunit, TGT, catalytic subunit, TGT_HUMAN, TGUT, qtrt1, tRNA-guanine transglycosylase

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Sanger Sequencing - Human QTRT1 knockout HeLa cell line (AB265454)
  • Sanger seq

Unknown

Sanger Sequencing - Human QTRT1 knockout HeLa cell line (AB265454)

Allele-1 : 46 bp deletion in exon 1.

Sanger Sequencing - Human QTRT1 knockout HeLa cell line (AB265454)
  • Sanger seq

Unknown

Sanger Sequencing - Human QTRT1 knockout HeLa cell line (AB265454)

Allele-2 : 17 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, 17 bp deletion in exon 1 and 46 bp deletion in exon 1

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
QTRT1
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 TGT protein also known as tRNA-guanine transglycosylase functions mechanically by modifying transfer RNA (tRNA) molecules through the exchange of guanine at specific tRNA positions with preQ1 precursors. This enzymatic activity is essential in tRNA maturation. The enzyme has a molecular mass of about 43 kDa. TGT is expressed mainly in the cytoplasm of eukaryotic cells and shows a high level of conservation across different species.
Biological function summary

The TGT catalyzed guanine exchange on tRNA enhances the correct folding of tRNA and supports the accuracy of protein translation. The protein is a part of an RNA-modifying enzyme family and does not typically form complexes but acts independently on its tRNA substrates. Its activity ensures proper tRNA structure influencing protein synthesis efficiency and overall cellular protein homeostasis.

Pathways

TGT plays an important role in the tRNA modification pathway which is important for post-transcriptional RNA processing. It connects with pathways involved in protein synthesis and RNA stability. TGT's function is integrally linked with enzymes like tRNA synthetases which are responsible for charging tRNAs with their corresponding amino acids establishing a pivotal link in the genetic code translation process.

Alterations in TGT function or expression impact specific disorders such as mitochondrial myopathy which results from defects in tRNA modification leading to flawed mitochondrial protein synthesis. TGT also has connections to cancer biology where aberrant tRNA modifications by TGT might influence tumoral cell growth and response to stress. In these contexts the interactions between TGT and other proteins involved in RNA metabolism such as Dicer point to a broader influence on cellular regulatory mechanisms linked to disease states.

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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