Human DICER1 Knockout Cell Line-HCT116
Cat.No. : CSC-RT0055
Host Cell: HCT116 Target Gene: DICER1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0055 |
| Cell Line Information | HCT116 -DICER1 (-/-) is a cell line with a homozygous knockout of human DICER1 |
| Target Gene | DICER1 |
| Host Cell | HCT116 |
| Species | 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 |
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Background
Case Study
Applications
DICER1, formally known as "Dicer 1, ribonuclease III," is a key gene in the biogenesis of microRNA (miRNA) and small interfering RNA (siRNA). These small RNA molecules play a key role in the regulation of gene expression, usually by directing the cleavage or translational repression of target messenger RNA (mRNA). The action of DICER1 begins with the generation of precursor miRNA (pre-miRNA) and precursor siRNA (pre-siRNA) in the nucleus. These precursors are exported to the cytoplasm, where DICER1 cleaves them into mature, functional forms. Mature miRNA and siRNA form RNA-induced silencing complexes (RISCs) that direct these complexes to complementary mRNA transcripts, thereby regulating gene expression post-transcriptionally.
Mutations and dysregulation of the DICER1 gene are associated with a variety of diseases, especially various forms of cancer. Germline mutations in DICER1 cause DICER1 syndrome, an inherited cancer predisposition syndrome that results in a variety of tumors, including pleuropulmonary blastoma (PPB), cystic nephroma, and multinodular goiter. Somatic mutations in DICER1 have also been associated with other sporadic tumors, such as Wilms tumor and certain types of ovarian and lung cancer.
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)
The human DICER1 knockout cell line (HCT116) provides a versatile platform for a wide range of research applications in molecular biology, cancer research, and genetics. Here are some of the key applications:
Cancer Research: Mutations or dysregulation of DICER1 have been implicated in various cancers, making this cell line an excellent model to study mechanisms of tumorigenesis, identify potential biomarkers, and develop new therapeutic strategies.
Gene Function Studies: Knockout of the DICER1 gene allows researchers to study the specific role of DICER1 in gene silencing, RNA interference, and microRNA biogenesis. This helps to elucidate how DICER1 contributes to cellular processes and how its loss affects gene expression profiles.
Drug Screening: The DICER1 knockout HCT116 cell line provides a unique platform for high-throughput drug screening. Researchers can test the efficacy and mechanism of new drugs targeting pathways affected by DICER1 loss, thereby facilitating the development of DICER1-specific or -related therapeutics.
Signaling Pathways: Researchers use this knockout cell line to study changes in various signaling pathways affected by DICER1 loss.
Developmental Biology: Because DICER1 is essential for microRNA processing, its knockout allows scientists to study developmental processes affected by microRNAs. This could improve the understanding of developmental disorders associated with DICER1 or microRNA dysfunction.
Synthetic Biology and Genetic Engineering: The DICER1 knockout HCT116 cell line can be used in synthetic biology to engineer cells with desired properties.
For research use only. Not intended for any clinical use.
