HCT116 DKO人结肠直肠癌基因双敲除细胞株NTCC® human colorectal cell line-BioVector NTCC典型培养物保藏中心

NTCC's HCT116 DKO cell line is a genetically modified version of the HCT116 human colorectal cancer cell line. The acronym "DKO" stands for "Double Knockout," indicating that two specific genes have been intentionally disabled. This cell line is a crucial tool in epigenetics research, particularly for studying the role of DNA methylation.

Origin and Genetic Modification

• Parental Cell Line: The DKO cell line originates from the HCT116 cell line, an adherent cell line derived from a human colon cancer. The wild-type HCT116 cell line is widely used in cancer research and is known for its near-diploid karyotype and stable genetic profile.

• Genetic Disruption: The key modification in the HCT116 DKO cell line is the targeted deletion of two genes:

  • $DNMT1$ (DNA methyltransferase 1): This enzyme is primarily responsible for maintaining existing methylation patterns during DNA replication.

  • $DNMT3B$ (DNA methyltransferase 3B): This enzyme is a de novo methyltransferase, meaning it is involved in establishing new methylation patterns.

• Resulting Phenotype: The simultaneous knockout of both DNMT1 and DNMT3B in the HCT116 DKO cells leads to a dramatic reduction in global DNA methylation. The methylation level is typically reported to be extremely low, often less than 5% of the wild-type HCT116 cells.

Research Applications

The HCT116 DKO cell line is a powerful model for researchers investigating the role of DNA methylation in gene regulation, cancer, and other biological processes. Its primary applications include:

• Epigenetics Research: It serves as an essential negative (unmethylated) control for various DNA methylation analysis methods, such as:

  • Bisulfite sequencing: This technique determines the methylation status of individual cytosine residues.

  • Methylation-specific PCR (MSP): A method used to detect methylation at specific CpG sites.

  • COBRA (Combined Bisulfite Restriction Analysis): A technique that uses restriction enzymes to assess methylation.

• Study of Epigenetically Silenced Genes: By comparing the gene expression profiles of wild-type HCT116 cells (which have many hypermethylated, silenced genes) with DKO cells, researchers can identify genes whose silencing is dependent on DNA methylation. Re-expression of previously silenced tumor suppressor genes in the DKO cells provides direct evidence of the role of methylation in their transcriptional control.

• Investigating the Functions of DNMTs: The DKO cells are used to understand the cooperative functions of DNMT1 and DNMT3B in maintaining DNA methylation patterns. Studies have shown that while single knockouts have minimal effects on methylation, the double knockout results in widespread demethylation.

• Cancer Biology: The DKO cell line helps in understanding how aberrant DNA methylation contributes to the transformed phenotype of cancer cells. The massive demethylation in DKO cells can lead to a less aggressive phenotype and increased apoptosis compared to the parental cell line.

Comparison with HCT116 Wild-Type

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