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Stable reporter cell lines are essential tools for monitoring gene expression, protein localization, and intracellular signaling in real time. Traditional approaches relying on random overexpression often suffer from position effects, variable expression levels, and potential interference with endogenous gene function. With the development of CRISPR/Cas9 technology, precise knock-in (KI) strategies have become the gold standard for reporter gene integration. By targeting specific genomic loci, reporter genes can be inserted into promoters or coding regions while preserving endogenous regulation, enabling stable, functional, and reproducible monitoring of cellular processes.
At Creative Biogene, our reporter genes knock-in platform combines years of expertise in genome editing and cell engineering. We offer high-efficiency integration of fluorescent proteins, luciferases, and epitope tags into safe-harbor loci such as Rosa26, AAVS1, and HPRT, or directly into target gene loci. This ensures long-term stable expression without affecting host cell proliferation, differentiation, or physiological functions.
Creative Biogene provides end-to-end services, from host cell selection to final delivery of fully validated reporter cell lines. Our standardized workflow is designed to maximize efficiency, accuracy, and reproducibility across diverse cell types:
1Design & Evaluation: Tailored knock-in strategies designed around your target gene and integration site, ensuring stable, high-value expression from the start.
2Construction & Editing: Advanced CRISPR/Cas9 and HDR workflows deliver precise, efficient integrations that save time and maximize success.
3Clone Screening & Validation: Rigorous verification by PCR, sequencing, and imaging guarantees accuracy, functionality, and confidence in your results.
4Functional Assays: Comprehensive testing of promoter activity, protein localization, and pathway activation ensures biological relevance and reproducibility.
5Final Delivery: Ready-to-use, fully validated cell stocks with complete QC data provide a seamless transition to downstream applications.
This streamlined workflow allows for both single-reporter and multi-reporter constructs, supporting complex pathway analysis and multi-gene dynamic studies.
Reporter knock-in cell lines are widely applied in functional genomics, signal transduction research, drug discovery, and in vivo imaging. Depending on the experimental goal, Creative Biogene can construct cell lines tailored to specific applications:
| Application | Reporter Type | Example Use |
| Gene Expression Analysis | Luciferase, GFP | Monitor promoter activity, transcription factor regulation, and complex gene networks |
| Protein Localization & Dynamics | GFP, RFP | Track subcellular protein distribution and dynamic changes in live cells |
| Signal Pathway Study | NF-κB/Luc, AP-1/Luc | Investigate pathway activation, receptor-ligand interactions, and cellular responses |
| Drug Screening & Mechanism Studies | Multi-reporter constructs | High-throughput compound screening, target validation, and safety assessment |
| In Vivo Imaging | Luc/EGFP | Monitor tumor growth, metastasis, or cell transplantation in live animal models |
Creative Biogene's reporter knock-in technology addresses common research challenges, ensuring reproducible results and reliable experimental models:
Our proprietary uniform knock-in process combines optimized sgRNA design, HDR template construction, and high-efficiency transfection strategies, enabling stable knock-in even in hard-to-transfect cells.
Creative Biogene provides clear guidelines to ensure successful cell line generation:
Deliverables include:
1. Knock-in Red Fluorescent Protein (RFP) gene into HEK293 cell line at the Rosa26 safe harbour site.
2. PCR detection of the genomes of transfected cells.
3. Sequencing results of the Rosa26 locus in RFP-positive HEK293 cells
Sequencing results of the upstream region of Rosa26 site.
Creative Biogene provides a complete, reliable, and customizable reporter knock-in cell line generation service. Our platform ensures precision, stability, and reproducibility, helping researchers advance functional genomics studies, drug discovery, and in vivo imaging projects. Contact Creative Biogene today to design your stable reporter knock-in cell line and accelerate your research.
Q: What types of reporters can be integrated?
A: We support fluorescent proteins (GFP, RFP), luciferases, and epitope tags (His, Flag), suitable for monitoring gene expression, protein localization, signaling pathways, and in vivo imaging.
Q: Which cell types are compatible?
A: HEK293, CHO, HeLa, human/mouse ESCs, primary cells, and many other mammalian cell types.
Q: How long does the process take?
A: Turnaround depends on cell type and reporter complexity, but our optimized workflow ensures faster delivery compared to traditional methods.
Q: Can multi-reporter cell lines be constructed?
A: Yes, we support single- or multi-reporter constructs for complex pathway analysis and dynamic studies.
Q: Is functional validation included?
A: Absolutely. Each cell line undergoes rigorous validation, including PCR, sequencing, imaging, and functional assays to ensure reliability.
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