Cat.No. : CSC-RT0055
Host Cell: HCT116 Target Gene: DICER1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
| Cat. No. | CSC-RT0055 |
| Description | HCT116 -DICER1 (-/-) is a cell line with a homozygous knockout of human DICER1 |
| Target Gene | DICER1 |
| Host Cell | HCT116 |
| Host Cell Species | Homo sapiens (Human) |
| 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 |
Emerging evidence suggests that microRNAs, a class of small and conserved noncoding RNAs, are involved in many physiological and pathological processes. The RNase III endonuclease DICER1 (DICER) is one of the key enzymes in microRNA biogenesis. Here, researchers found that DICER was downregulated at both the mRNA and protein levels in tumor samples from patients with colorectal cancer (CRC). Importantly, mice with intestinal epithelial cell (IEC)-specific deletion of Dicer developed increased tumors after administration of azoxymethane and dextran sodium sulfate (DSS). Microarray analysis of 3 paired Dicer deletion CRC cell lines showed that miR-324-5p was one of the most significantly decreased miRNAs. Significant reduction of miR-324-5p was also found in the intestinal epithelium of mice with IEC-specific deletion of Dicer. Mechanistically, miR-324-5p directly binds to the 3′ untranslated region (3′UTR) of HMG-box 3 (HMGXB3) and WAS protein family member 2 (WASF-2), thereby inhibiting their expression. These two proteins are key proteins involved in cell motility and cytoskeletal remodeling. The studies here reveal that the DICER/miR-324-5p/HMGXB3/WASF-2 axis plays a key role in CRC tumorigenesis by regulating cytoskeletal remodeling and maintaining the integrity of the intestinal barrier.
Human MiRNA microarray analysis was performed using three pairs of DICER-WT and DICER knockout CRC cell lines (RKO, HCT116, and DLD) to identify key miRNAs affected by DICER deficiency. hsa-miR-324-5p was one of the two microRNAs that decreased most significantly after DICER deficiency compared to wild-type cells, and has-miR-324-5p levels were significantly reduced in the three DICER-deficient cells (Figure 1A). RT-PCR experiments confirmed that has-miR-324-5p expression was reduced in all three DICER-deficient cell lines (Figure 1B). In mice with IECs specific Dicer deficiency (Dicerloxp/loxp&VillinCre and Dicerloxp/+&VillinCre), mmu-miR-324-5p mRNA levels were also significantly decreased in the intestine epithelial layer (Figure 1C). In addition, RT-PCR experiments showed that the level of has-miR-324-5p mRNA was also significantly suppressed after siDICER transfection in RKO (Figure 1D) and HCT116 cells (Figure 1E). Taken together, these data indicate that miR-324-5p is one of the key and indispensable DICER downstream microRNAs involved in cytoskeletal remodeling.
Figure 1. DICER deletion is accompanied with miR-324-5p downregulation. (Sun, Li Na, et al. 2017)
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.
A: Single clonal cell.
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.
A: The cell line should be stored in liquid nitrogen for long-term preservation.
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.
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
The complete knockout of DICER1 allows us to study gene silencing pathways more effectively. This has led to quicker discoveries and advancements, saving us both time and resources in our experiments.
The Human DICER1 Knockout Cell Line-HCT116 has significantly improved the precision of our genetic studies. It replicates the human physiological environment remarkably well, providing us with highly reliable data.
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