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Panoply™ Human PRAME Over-expressing Stable Cell Line

Panoply™ Human PRAME Over-expressing Stable Cell Line

Cat.No. :  CSC-SC012314 Host Cell:  HEK293 (CHO and other cell types are also available)

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Gene Informationn

Cat. No. CSC-SC012314
Description Using Creative Biogene's proprietary lentiviral vectors, we subclone the target gene into lentivector, generate the lentivirus particles, sequentially infect the cell line HEK293 (other cell types are also available according to your requirements), and select the clones constantly expressing target gene at high level.
Gene PRAME
Gene Species Homo sapiens (Human)
Host Cell HEK293 (CHO and other cell types are also available)
Stability Validated for at least 10 passages
Application

1. Gene expression studies

2. Signaling pathway research

3. Drug screening and toxicology

4. Disease research

Quality Control Negative for bacteria, yeast, fungi and mycoplasma.
Size Form 2 × 10^6 cells / vial
Shipping Dry Ice
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.
Gene Name
Gene Symbol
Synonyms
Gene Description
Gene ID
UniProt ID
mRNA Refseq
Protein Refseq
Chromosome Location
Function
MIM
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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Preferentially expressed antigen (PRAME) in melanoma belongs to the CTA gene family and is aberrantly expressed in various cancer types. Previous studies have found that PRAME is highly expressed in cervical cancer tissues but not in normal tissues. Here, researchers examined PRAME expression in cervical tissues and cells using immunohistochemistry (IHC), qRT-PCR, and Western blotting. CCK-8, BrdU, wound healing assays, Transwell assays, and flow cytometry were used to investigate the role of PRAME in regulating the malignant biological behavior of cervical cancer cells. IHC, qRT-PCR, and Western blotting results showed that PRAME is highly expressed in cervical cancer tissues and cells. Knockdown of PRAME attenuated the growth, migration, and invasion of C33A and SiHa cells, induced G0/G1 arrest, and increased apoptosis by regulating the Wnt/β-catenin signaling pathway. However, upregulation of PRAME exhibited the opposite effect, which was partially reversed by MSAB treatment. When PRAME is overexpressed by activating the Wnt/β-catenin signaling pathway, the growth rate of xenograft tumors is enhanced. In summary, PRAME is associated with the development and progression of cervical cancer mediated by the Wnt/β-catenin signaling pathway, suggesting that PRAME may be one of the factors regulating the development of cervical cancer and a potential therapeutic target.

The cell cycle distribution of PRAME-knockdown and PRAME-overexpressing cells was assessed with PI staining by using flow cytometry, and cell cycle profiles were fitted by Flowjo software. As shown in Figures 1A and 1B, PRAME knockdown cells were more likely to arrest in the G0/G1 phase than corresponding control cells, while the proportion of cells in the S phase was lower in the PRAME knockdown group than in the control group. Compared with the control group, PRAME overexpression reduced the arrest of cells in the G0/G1 phase in C33A and SiHa cells, while the proportion of cells in the S phase in PRAME-overexpressing C33A and SiHa cells was higher than in the control group (Figures 1C and 1D). This suggests that PRAME knockdown disrupts cell cycle progression, while PRAME overexpression reduces the arrest of cells in the G0/G1 phase.

Figure 1. The effect of PRAME expression on cell cycle distribution of C33A and SiHa cells.Figure 1. The effect of PRAME expression on cell cycle distribution of C33A and SiHa cells. (Chen X, et al., 2023)

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