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  The 4T1 Luciferase (4T1-Luc) stable cell line is a mouse breast cancer cell line engineered to stably express the Firefly Luciferase reporter gene, derived from the highly metastatic parental 4T1 line. Through bioluminescence imaging, the 4T1-Luc cell line enables real-time, non-invasive monitoring of tumor growth and metastasis, making it an ideal model for studies on breast cancer metastasis, drug screening, and immunotherapy.
  This instruction provides a detailed overview of how to use the 4T1-Luc cell line to establish breast cancer metastasis models and apply them in scientific research, covering experimental protocols and optimization strategies. By using our ready-to-use 4T1-Luc product, researchers can rapidly initiate experiments and significantly reduce the time required for model construction.
  

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     Advantages of the 4T1-Luc Cell Line in Breast Cancer Research
  

  The 4T1-Luc cell line, with its high metastatic potential and stable bioluminescent signal, has become a key tool in breast cancer research and is suitable for the following applications:

  • ● Metastasis studies: Models lung, liver, and bone metastasis of triple-negative breast cancer, allowing real-time tracking of metastatic progression.
  • ● Drug screening: High-sensitivity bioluminescence enables high-throughput screening of chemotherapeutics, targeted therapies, or nanomedicines.
  • ● Immunotherapy research: Used in combination with CAR-T cells or PD-1 inhibitors to evaluate changes in the tumor immune microenvironment.
  • ● Gene function validation: Supports gene knockout or overexpression (e.g., CRISPR-mediated E-cadherin editing) to study EMT pathways.
  • ● In vivo and ex vivo modeling: Compatible with orthotopic mouse models; non-invasive IVIS imaging reduces the number of animals required.

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    Product Recommendation
  

  The 4T1-Luc cell line provided by EDITGENE exhibits >99% purity and high activity, with bulk orders eligible for discounts.

  Luciferase Assay Results:

  Sample	Replicate 1	Replicate 2	Replicate 3	Mean	Fold Expression
  WT	1550	2368	4094	2671	42
  LUC	107153	113540	112395	111029

  
  Advantages:

  ● Purity >99%, activity >1.2 × 10⁶ RLU/10⁵ cells (n=3, SD <5%).
  ● Stable for >20 passages; metastasis efficiency >80% (validated by IVIS).
  ● COA and SOP are provided; discounts available for bulk orders.

Pinciple of Construction

The 4T1-Luc cell line is engineered using genetic modification techniques, where the luciferase (luc) gene is inserted into a suitable vector and transfected into the parental 4T1 cells to achieve stable expression. Luciferase catalyzes the oxidation of its substrate, luciferin, generating bioluminescence with a peak wavelength of approximately 550-570 nm. The luminescence intensity correlates linearly with enzyme expression levels. Common configurations include single-luciferase systems or dual-luciferase systems, the latter incorporating Renilla luciferase as an internal control for experimental normalization.

Common Construction Methods

The most widely used approach is lentiviral transduction, as it allows efficient and stable integration. The following outlines a typical workflow using EDITGENE’s 4T1-Luc cell line as an example:

1. Construct the expression vector:

• a. Clone the luciferase (luc) gene into a lentiviral vector (e.g., pLenti or pLVX).
• b. The vector typically includes a promoter (such as CMV) to drive luc expression, a selectable marker (e.g., puromycin resistance) for screening, and a fluorescent tag (e.g., GFP) for visualization.
• c. For dual-luciferase systems, Renilla luciferase can be included as an internal control, either co-expressed or placed on the same plasmid (e.g., pGL3 or pmirGLO).

  
2.Lentivirus packaging:

• a. Co-transfect the recombinant vector with helper plasmids (packaging plasmid psPAX2 and envelope plasmid pMD2.G) into HEK293T cells to produce lentiviral particles.
• b. Collect the viral supernatant, filter, and determine titer (typically 10⁸-10⁹ TU/mL)

  
3. Transduce parental cells:

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  • 4. Select stable clones:
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• • （3D single-cell printing precisely separates individual cells, greatly improving the accuracy of monoclonal screening and cell survival. This non-contact method avoids mechanical damage and background contamination, maintaining cell integrity and biological activity. Compared with traditional limiting dilution, 3D single-cell printing reduces human error and ensures reliable screening outcomes.）
  
  
  • 5. Validation and Applications
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  • a. Validation Methods: Gene integration can be confirmed by qPCR, protein expression by Western blot, and luciferase activity by bioluminescence assay.
  • b. Validation Criteria: Luciferase activity > 10⁶ RLU/10⁵ cells, with stable expression maintained for over 20 passages.
  • c. Applications: Cells can be injected into mouse models, and tumor growth can be monitored using an IVIS imaging system after intraperitoneal administration of the luciferase substrate.ction.
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    Application Examples: Advancing Research Projects
  
  • 4.1  Breast Cancer Metastasis Mechanism