Cat.No. : CSC-RR0210 Host Cell: HT-29
| Cat. No. | CSC-RR0210 |
| Description | HT29-GFP/Luc is derived from HT29 cell line which is engineered by lentivirus transduction to enable stable expression of EGFP and firefly luciferase. This cell line is an ideal positive control for use in both fluorescence and bioluminescence assays. |
| Background | The Luc/GFP reporter cell line HT-29 has emerged as a vital tool in molecular biology research |
| Introduction | Luc/GFP Reporter Cell Line-HT-29 is a polyclonal population which is constructed by lentivirus transduction, followed by stable cell selection. This cell line stably expresses luciferase gene under the control of CMV promoter. It also stably expresses GFP and puromycin resistance gene driven by SV40 promoter. This cell line is designed for in vitro or in vivo cancer research. |
| Gene | GFP/Luc |
| Host Cell | HT-29 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Reporter Type | Fluorescent protein |
| Application | 1. Gene expression studies 2. Protein localization 3. Drug screening and toxicology 4. Live cell imaging |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Source | HT-29 |
| 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 |
ALDHs are NAD(P)+-dependent enzymes converting aldehydes to carboxylic acids. Researchers investigated the role of ALDH1B1 in human colorectal adenocarcinoma using Luc/GFP Reporter Cell Line-HT-29. They stably transfected HT29 cells with GFP-tagged ALDH1B1 or an empty lentiviral vector. Overexpression of ALDH1B1 led to altered cell morphology, decreased proliferation, and reduced clonogenicity. ALDH1B1 induced G2/M arrest, possibly via p53 and p21 upregulation. Moreover, ALDH1B1-overexpressing HT29 cells showed resistance to doxorubicin, 5-FU, and etoposide, along with enhanced migratory potential and epithelial-mesenchymal transition (EMT) markers upregulation. These findings underscore ALDH1B1's potential role in cancer progression by influencing cell morphology, gene expression, chemosensitivity, and migratory capabilities.
Figure 1. The HT29/ALDH1B1 cell line, created by stable transfection of human ALDH1B1 cDNA into HT29 cells, exhibits over 17-fold higher ALDH1B1 expression than HT29/mock cells. Flow cytometry confirms GFP-tagged ALDH1B1 expression, with 92.9% GFP+ in HT29/ALDH1B1 versus 96.1% GFP− in HT29/mock. Regular monitoring ensures stable ALDH1B1 expression. Enhanced GFP fluorescence in HT29/ALDH1B1 cells is evident compared to HT29/mock cells via fluorescence microscopy. (Tsochantaridis I, et al., 2021)
A: HT-29 cells were chosen for establishing the luciferase/GFP reporter cell line due to their relevance to colorectal cancer research and their ability to serve as a model system for studying gene expression and signaling pathways. Additionally, HT-29 cells are well-characterized and widely used in cancer biology studies.
A: The stability and expression level of luciferase and GFP in the HT-29 reporter cell line were verified and maintained through stable transfection, antibiotic selection, and validation of expression using luciferase assays and fluorescence microscopy. Continuous culture under appropriate conditions ensured sustained expression levels.
A: Functional characterization of luciferase and GFP expression in the HT-29 reporter cell line involved assessing their responsiveness to regulatory elements by analyzing changes in luciferase activity and fluorescence intensity in response to stimuli. Additionally, their involvement in cellular processes such as gene regulation, signaling pathways, or drug response was investigated using appropriate assays and imaging techniques.
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An invaluable resource! The Luc/GFP Reporter Cell Line has significantly enhanced my research capabilities, offering a versatile platform for studying cancer biology and evaluating therapeutic interventions in vitro. Its dual reporter expression simplifies experimental workflows, allowing for efficient data collection and analysis, and accelerating discoveries in cancer research.
Reliable dual expression! This cell line surpasses expectations, providing both luciferase bioluminescence and GFP fluorescence, facilitating comprehensive and quantitative analysis of cellular events.
The Luc/GFP Reporter Cell Line in HT-29 cells offers dual expression of luciferase and GFP, enabling multifaceted analysis of cellular processes in my cancer research.
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