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Human ABCB4 Knockout Cell Line-Huh7

Human ABCB4 Knockout Cell Line-Huh7

Cat.No. :  CSC-RT2766

Host Cell:  Huh7 Target Gene:  ABCB4

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

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2766
Description This cell is a stable cell line with a homozygous knockout of human ABCB4 using CRISPR/Cas9.
Target Gene ABCB4
Host Cell Huh7
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 ABCB4 gene, also known as MDR3, is an important part of the human genome and is responsible for encoding a member of the ATP-binding cassette (ABC) transporter proteins. This particular protein plays a crucial role in the movement of phospholipids across cell membranes, specifically from liver cells into bile. Phospholipids are key molecules that stabilize bile acids, making them less toxic to cells and therefore essential for healthy bile composition. When ABCB4 functions properly, it ensures proper emulsification of dietary fats and protects liver cells from the detergent-like effects of bile acids. Mutations in the ABCB4 gene have been linked to a variety of diseases, including progressive familial intrahepatic cholestasis type 3 (PFIC3), a severe disease that manifests in early childhood and can lead to liver failure. Variants in this gene disrupt the normal transport of phospholipids, leading to a buildup of toxic free bile acids in the liver, which in turn causes liver cell damage and cholestasis. Another important disease associated with ABCB4 mutations is intrahepatic cholestasis of pregnancy (ICP), a liver disease characterized by severe itching in the second half of pregnancy. The disease is caused by a genetic predisposition, with affected individuals having variants that result in less efficient phospholipid transport. The increased metabolic burden during pregnancy exacerbates this problem, leading to a buildup of bile acids that impair liver function and cause cholestatic symptoms.
The human ABCB4 knockout cell line - Huh7 was designed for a variety of applications in the biomedical research field, providing important insights into liver biology and related diseases. Here are some of the applications of this cell line: Lipid Metabolism Studies: The ABCB4 gene plays a key role in lipid trafficking in hepatocytes. Using the ABCB4 knockout Huh7 cell line, researchers can study disruptions in lipid metabolism and their consequent effects, which are critical to understanding diseases such as cholestasis and liver fibrosis. Drug Testing and Toxicology: Huh7 cells lacking ABCB4 can be used to evaluate the hepatotoxicity of new drugs. By observing how these drugs affect hepatocytes lacking a functional ABCB4 gene, researchers can determine potential adverse effects and mechanisms of drug-induced liver injury. Cholesterol Regulation Studies: ABCB4 is also involved in the secretion of cholesterol into bile. This knockout model can be used to study pathways that regulate cholesterol levels in the liver and help identify therapeutic targets for hypercholesterolemia and related diseases. Gene Therapy Studies: This cell line can serve as a model to evaluate the efficacy of gene therapy techniques designed to correct ABCB4 defects. Researchers can test the introduction of a functional ABCB4 gene or its modifications to restore normal cellular function and pave the way for therapeutic intervention. Disease Modeling: The ABCB4 knockout Huh7 cell line is particularly valuable in modeling liver diseases such as progressive familial intrahepatic cholestasis (PFIC). This model mimics the genetic and phenotypic features of these diseases, aiding the study of their progression and the development of potential treatments.
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
The cell line's quality is top-notch!

The cell line's quality is top-notch! The cells arrived in excellent condition, with high viability and optimal confluency.

Germany

01/05/2023

Very helpful

Utilizing the Human ABCB4 Knockout Cell Line-Huh7 has significantly accelerated our drug discovery process. The knockout model mimics clinical conditions more accurately, enabling us to understand drug interactions and mechanisms of resistance better.

United States

07/27/2021

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