Cas9 Stable Cell Line - BXPC-3
Cat.No. : CSC-RO0032 Host Cell: BXPC-3
Size: >1x10^6 cells/vial Validation: T7 Endonuclease I assay
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Cell Line Information
Safety and Packaging
| Cat. No. | CSC-RO0032 |
| Product Type | Cas9 overexpression stable cell line |
| Introduction | Clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 is a gene-editing technology that contains two essential components: a guide RNA (gRNA) to match a target gene, and the Cas9 (CRISPR-associated protein 9) endonuclease which causes a double-stranded DNA break, allowing modifications to the genome via nonhomologous end joining (NHEJ) or homology-directed repair (HDR). |
| Cell Line Information | BXPC-3-Cas9 cell line is engineered to stably overexpress Cas9 nuclease. The Cas9 nuclease in BXPC-3-Cas9 cell line has been functionally validated using T7 Endonuclease I assay. In combination with separately transfected sgRNAs, BXPC-3-Cas9 cell line can be used to efficiently generate targeted genomic modifications including gene knockout, gene knockin, gene mutagenesis, gene tagging etc. It is also an ideal cell line model for sgRNA screening and validation, either individually or in pools. |
| Target Gene | Cas9 |
| Host Cell | BXPC-3 |
| Applications | 1) CRISPR genome editing, such as gene knockout (KO), gene knockin (KI), gene mutagenesis, gene tagging etc. 2) High-throughput sgRNA screening and validation |
| Quality Control | 1) T7E1 assay 2) Mycoplasma detection |
| Size Form | One vial of frozen cells, typically >1x10^6 cells/vial |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| 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
Cas9, a pivotal RNA-guided DNA endonuclease, emerged as a groundbreaking discovery in 2012 within the framework of the bacterial CRISPR-Cas system. Its profound impact on molecular biology cannot be overstated. Simultaneously, BXPC-3, a human pancreatic cancer cell line, has played a crucial role in cancer research owing to its physiological relevance and genetic characteristics. The fusion of these two entities resulted in the creation of the Cas9 Stable Cell Line-BXPC-3. This milestone achievement, likely realized in the subsequent years following Cas9's discovery, marks a significant advancement in cancer biology. By integrating the Cas9 gene into the BXPC-3 genome, researchers gained a powerful tool for precise genetic manipulation within the context of pancreatic cancer.
The establishment of the Cas9 Stable Cell Line-BXPC-3 facilitates targeted genetic modifications, providing invaluable insights into the underlying mechanisms of pancreatic cancer progression and response to therapeutic interventions. This innovative cell line exemplifies the ongoing refinement and utilization of CRISPR-Cas9 technology in disease modeling and the development of targeted therapies, underscoring its transformative potential in advancing biomedical research and clinical applications.
TGF-β signaling is essential in pancreatic ductal adenocarcinoma (PDAC) development, altered in all PDAC cases as one of four major pathways. Researchers demonstrate the crucial role of SMAD2/3 in pancreatic ductal adenocarcinoma (PDAC) progression, particularly in SMAD4-null contexts. By investigating TGF-β signaling in PDAC cells lacking SMAD4, they reveal SMAD2/3's oncogenic effects, influencing collective migration through FAK and Rho/Rac signaling pathways. RNA-sequencing analyses unveil a TGF-β gene signature linked to aggressive behavior, facilitated by SMAD2/3 activation. Additionally, clinical data suggests that SMAD4-negative tumors with elevated phospho-SMAD2 levels exhibit increased aggressiveness and poorer prognosis. These findings underscore the complex interplay between TGF-β signaling components in PDAC, highlighting SMAD2/3 as potential therapeutic targets.
Figure 1. A double SMAD2/SMAD3 knockout BxPC-3 (SMAD4-negative) pancreatic cancer cell line was generated. SMAD protein expression and localization in response to TGF-β treatment were assessed. Altered SMAD2/3 levels and distribution were observed in knockout cells compared to controls, providing insights into TGF-β signaling in pancreatic cancer progression. (Bertrand-Chapel A, et al., 2022)
Utilizing Creative Biogene's Cas9 Stable Cell Line-BXPC-3 streamlines the process of SMAD2/SMAD3 gene knockout experiments. This stable cell line is directly amenable to CRISPR/Cas9 gene editing technology, facilitating the rapid generation of SMAD2/SMAD3 knockout cell lines, thereby saving time and resources. Such cell lines can be instrumental in investigating the TGF-β signaling pathway and its implications in pancreatic cancer progression, enhancing the convenience and efficiency of related disease research endeavors.
1. Gene Editing: Utilizing Cas9 Stable Cell Line-BXPC-3 for targeted gene knockout studies in pancreatic cancer research.
2. Drug Screening: Employing Cas9 Stable Cell Line-BXPC-3 to assess drug efficacy and resistance mechanisms in pancreatic cancer cells.
3. Functional Genomics: Investigating the role of specific genes in pancreatic cancer progression using Cas9 Stable Cell Line-BXPC-3.
4. Therapeutic Development: Validating potential therapeutic targets through CRISPR/Cas9-mediated gene editing in BXPC-3 cells.
5. Metastasis Study: Examining the impact of gene mutations on pancreatic cancer metastasis using Cas9 Stable Cell Line-BXPC-3.
6. Immunotherapy Evaluation: Assessing immune checkpoint inhibitors' effectiveness in BXPC-3 cells modified with Cas9 technology.
7. Tumor Microenvironment Research: Exploring interactions between pancreatic cancer cells and their microenvironment using Cas9 Stable Cell Line-BXPC-3.
8. Cancer Stem Cell Investigation: Characterizing cancer stem cells in pancreatic cancer using Cas9 Stable Cell Line-BXPC-3.
For research use only. Not intended for any clinical use.
