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AB265243

Human PATL1 (Pat1b) knockout HeLa cell line

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PATL1 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 6 and Insertion of the selection cassette in exon 6.

View Alternative Names

OK/KNS-cl.5, PAT1-like protein 1, PATL1_HUMAN, Pat1b, Protein PAT1 homolog 1, Protein PAT1 homolog b, hPat1b, protein associated with topoisomerase II homolog 1

2 Images
Sanger Sequencing - Human PATL1 (Pat1b) knockout HeLa cell line (AB265243)
  • Sanger seq

Unknown

Sanger Sequencing - Human PATL1 (Pat1b) knockout HeLa cell line (AB265243)

Allele-2 : Insertion of the selection cassette in exon 6.

Sanger Sequencing - Human PATL1 (Pat1b) knockout HeLa cell line (AB265243)
  • Sanger seq

Unknown

Sanger Sequencing - Human PATL1 (Pat1b) knockout HeLa cell line (AB265243)

Allele-1 : 1 bp insertion in exon 6.

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 insertion in exon 6 and Insertion of the selection cassette in exon 6

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, 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
PATL1
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.

Pat1b also known as Processing body component 4 (PATI-4) is a protein with a mass of approximately 73 kDa. It functions mechanically as an RNA-binding protein involved in mRNA decay. Pat1b enhances mRNA decay by promoting decapping and subsequently exonucleolytic degradation. It is expressed in various human tissues with prominent activity reported in the brain and testes. In molecular biology labs Pat1b is regularly studied due to its central role in mRNA turnover.
Biological function summary

The degradation of mRNA represents a critical step where Pat1b plays an essential role. As a component of the Pat1-Lsm complex Pat1b interacts with several proteins to regulate the stability and translation of mRNA. Pat1b influences the mRNA decapping process which is vital for controlled mRNA degradation thereby controlling gene expression post-transcriptionally. This process ensures the removal of aberrant or unnecessary mRNAs from the cellular environment enabling the cell to adapt to new conditions or signals rapidly.

Pathways

MRNA decay and processing pathways involve Pat1b as a central component. Within the mRNA surveillance pathway Pat1b cooperates with Dcp1-Dcp2 decapping enzymes to mediate mRNA decay. Pat1b is pivotal in the nonsense-mediated mRNA decay (NMD) pathway where it partners with Upf1 to target and degrade mRNAs containing premature stop codons. These pathways ensure cellular integrity and protein synthesis regulation.

Disruptions in Pat1b are linked to neurodegenerative diseases such as Amyotrophic lateral sclerosis (ALS) and conditions like intellectual disability. In ALS a failure in precise mRNA surveillance and decay pathways leads to toxic buildup of improperly processed mRNAs implicating Pat1b along with proteins such as TDP-43. In the context of intellectual disabilities altered mRNA metabolism involving Pat1b and its interaction with proteins like FMRP can affect neuronal function and development highlighting its significance for normal cognitive function.

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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