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Human WRN Knockout Cell Line-HCT116

Human WRN Knockout Cell Line-HCT116

Cat.No. :  CSC-RT2776

Host Cell:  HCT116 Target Gene:  WRN

Size:  1x10^6 cells/vial, 1mL Validation:  Sequencing

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2776
Description This cell is a stable cell line with a homozygous knockout of human WRN using CRISPR/Cas9.
Target Gene WRN
Host Cell HCT116
Host Cell Species Homo sapiens (Human)
Size Form 1 vial (>10^6 cell/vial)
Shipping Dry ice package
Storage Liquid Nitrogen
Revival Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media.
Mycoplasma Negative
Format One frozen vial containing millions of cells
Storage Liquid nitrogen
Safety Considerations

The following safety precautions should be observed.

1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.

2. No eating, drinking or smoking while handling the stable line.

3. Wash hands after handling the stable line and before leaving the lab.

4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.

5. All waste should be considered hazardous.

6. Dispose of all liquid waste after each experiment and treat with bleach.

Ship Dry ice
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Background

Applications

Publications

Q & A

Customer Reviews

The WRN gene encodes the Werner protein, an important enzyme with both helicase and exonuclease activities. The main function of the helicase is to unwind and separate double-stranded DNA, while the exonuclease trims the broken ends of damaged DNA by removing nucleotides. The Werner protein first unwinds the DNA and then removes abnormal DNA structures that may have formed unintentionally. Both steps are essential for maintaining the structural integrity of an individual's DNA. In addition to its role in DNA repair, the Werner protein is also essential in the processes of DNA replication and transcription. Mutations in the WRN gene are directly associated with Werner syndrome, a disorder characterized by premature aging and increased susceptibility to various age-related diseases, including cancer. More than 60 different mutations have been identified, and most result in a truncated and nonfunctional Werner protein. This defective protein fails to localize to the nucleus (where it normally interacts with DNA) and is often degraded more rapidly than the functional protein. The lack of sufficient Werner protein in the nucleus impairs the processes of DNA replication, repair, and transcription. In addition to the inherited mutations that cause Werner syndrome, somatic changes in the WRN gene can also lead to cancer. These acquired changes may involve hypermethylation, the process of adding methyl groups to DNA, which effectively silences the WRN gene. This gene silencing prevents the production of the Werner protein, which leads to the accumulation of mutations and can cause uncontrolled cell growth and tumor formation. This hypermethylation has been observed in a variety of tumors, including those of the colon, rectum, lung, stomach, prostate, breast, and thyroid.
The WRN (Werner syndrome, RecQ helicase-like) gene plays an important role in maintaining genome stability through DNA repair and replication processes. The knockout cell line of WRN in HCT116 cells offers numerous research applications. The following are the main applications of this cell line: 1. Cancer Research: The WRN knockout cell line helps study the mechanisms of tumorigenesis. WRN loss-of-function mutations are associated with genomic instability and increased cancer risk, making this cell line valuable for understanding the genetic pathways that lead to cancer development. 2. Aging Research: Werner syndrome is characterized by premature aging due to WRN deficiency. The WRN knockout cell line provides a model to study the molecular mechanisms of aging and screen potential anti-aging therapies targeting pathways affected by WRN loss. 3. DNA Repair Mechanisms: The WRN protein participates in multiple DNA repair pathways such as base excision repair (BER), nucleotide excision repair (NER), and homologous recombination (HR). This cell line helps to elucidate the role of these pathways and the specific contribution of WRN to DNA damage repair and genome integrity. 4. Drug Screening: WRN knockout cells can be used to screen for compounds that compensate for WRN deficiency or enhance other DNA repair pathways. This is critical for developing drugs that target specific cancers with WRN mutations or defects. 5. Metabolic and Neurodegenerative Disease Research: Given WRN's role in maintaining telomere stability and its interaction with various cellular proteins, WRN knockout cell lines can be used to study its effects on metabolic diseases such as diabetes and neurodegenerative diseases such as Alzheimer's, where genome stability and DNA repair are impaired.
Customer Q&As
How is the knockout cell line validated?

A: The knockout cell product is validated by PCR amplification and Sanger Sequencing to confirm the mutation at the genomic level. Please find the detailed mutation info in the datasheet.

Is the product a single clonal cell or mixed cell pool?

A: Single clonal cell.

Can I confirm gene knockout by RT-qPCR?

A: No. This knockout cell product is generated using the CRISPR/Cas9 system to induce small insertions or deletions (indels) resulting in frameshift mutations. Although these frameshift mutations typically disrupt the coding gene, there is a possibility that the non-functional transcript may still be transcribed. Consequently, this could potentially yield misleading results when analyzed by RT-qPCR.

How can I store the cell product?

A: The cell line should be stored in liquid nitrogen for long-term preservation.

Is it possible to get multiple knockout clones for my GOI?

A: For most cases, we often keep at least 2 clones with different frameshift mutations. Please feel free to contact us to check if there are additional available clones.

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Customer Reviews
We achieved good experimental results.

WRN knockout cells can be used to identify potential therapeutic targets for Werner syndrome. We achieved good experimental results.

United States

11/03/2022

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