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Functional Pathway Analysis with CRISPR KO & TF Reporter System

Signosis offers over 80 stable TF reporter cell lines, including reporters for NRF2, NFκB, p53, SMAD1/5/8, and others, alongside a comprehensive suite of CRISPR knockout plasmids targeting pathway regulators, kinases, and receptors.

Principle 

The Knockout-TF Reporter Combo System is a ready-to-use assay platform that integrates CRISPR-mediated gene knockout with our transcription factor (TF)-responsive luciferase reporter cell lines. Designed for pathway discovery, compound screening, and functional genomics, this system allows straightforward and sensitive luciferase readouts to easily monitor the effects of CRISPR knockouts.

Our CRISPR KO vector employs a dual sgRNA system optimized for single-gene knockout. Two independently expressed sgRNAs under U6 and H1 promoters target multiple regions within the same gene, ensuring precise and efficient disruption. This enhanced design increases knockout success and overall editing efficiency.

Researchers can directly combine our TF specific reporter lines with specific gene-targeted CRISPR knockout plasmids for a fast, sensitive way to study gene function and signaling pathways. CRISPR disrupts the target gene, while the reporter provides real-time, quantitative luciferase readouts of downstream TF activity.


Why use this system?

  • Direct functional readout: Confirm knockout effects by measuring TF activity changes.

  • Link genotype to phenotype: Quickly see how gene edits affect pathways.

  • Pathway-specific insights: Use pathway-targeted reporters (e.g., NFκB, STAT3) to uncover gene roles and crosstalk.

  • High-throughput ready: Fast, scalable luciferase assays replace complex methods.

  • Consistent results: Stable reporter lines ensure reproducibility.

  • Plug and Play: Seamlessly combine any KO Vector with our TF Reporter Cell Lines to uncover gene-pathway interactions—no cloning or optimization needed.

Example use cases:

  • NRF2 Reporter Cell Line + NRF2 Knockout: Decrease in luciferase signal under both basal and induced conditions.

  • NRF2 Reporter Cell Line + KEAP1 Knockout: Increased basal and induced luciferase activity due to constitutive activation of NRF2

Keap1–Nrf2 Pathway.png

Pathway Mapping and Deconvolution

Understanding how signaling pathways are regulated at the transcriptional level is critical for basic and translational research. By combining targeted CRISPR knockouts with transcription factor (TF) luciferase reporter cell lines, researchers can systematically interrogate the functional impact of individual genes on pathway activity. Knockouts of TFs (e.g., NRF2) or their regulators (e.g., KEAP1) allow users to distinguish upstream activators, repressors, and indirect modulators within complex signaling cascades.

Molecular Application
Slideshow

Product Pathways

All CRISPR Products

Signosis offers over 80 stable TF reporter cell lines, including reporters for NRF2, NFκB, p53, SMAD1/5/8, and others, alongside a comprehensive suite of CRISPR knockout plasmids targeting pathway regulators, kinases, and receptors. Browse our products below, or check out the specialized categories for each pathway in one of the tabs above. 

Benefits

Pathway Discovery, Simplified

Easily map signaling networks by pairing knockout plasmids with reporter lines for key transcription factors like NRF2, NFκB, p53, SMAD1/5/8, and more.

Precision-Driven Gene Functionality

Provides precise, gene-specific insights by linking CRISPR-induced knockouts to direct measurement of TF activity, revealing the impact of specific genes on cellular signaling.

Seamless Reporter Integration

Easily integrate TF reporters with gene knockouts to study how upstream genetic changes affect transcriptional outputs, enabling rapid mechanistic insights with minimal setup.

One Powerful System

Combines CRISPR knockout and TF-responsive reporters in one modular system to directly link gene disruption with transcriptional outcomes.

Accelerated Drug Mechanism Studies

Uncover molecular mechanisms behind drug efficacy or resistance by correlating knockout-induced changes in TF activity with drug treatment responses.

Precise Functional Genomics

Gain accurate, high-resolution insights into the functional roles of genes in transcriptional regulation, facilitating more targeted investigations in your research.

Ready-to-Use

Ready for immediate use, reducing setup time and enhancing experimental throughput in pathway discovery and functional genomics.

Flexible Experimental Design

Customize experiments, pairing different knockout plasmids with various TF reporters, making it ideal for diverse research goals, from basic biology to drug discovery.

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