High throughput dual-luciferase assays for promoter activity in protoplasts
High throughput dual-luciferase assays in protoplasts enable rapid, quantitative analysis of promoter variants. This technology facilitates the screening of single-nucleotide promoter changes, supporting efficient promoter engineering in plant species.
Overview
The high throughput dual-luciferase assay platform offers a robust method for analyzing promoter activity through the screening of single-nucleotide variants. By utilizing a dual-luciferase system, this technology allows for the rapid and quantitative assessment of changes in promoter activity. The platform is designed to handle a large library of promoter variants, facilitating efficient promoter engineering across various plant species. This method supports the discovery of promoter variants with enhanced or modified activity, crucial for advancements in plant biotechnology.
Technical specifications
- Dual-luciferase system: Utilizes firefly luciferase for measuring promoter activity and Renilla luciferase for normalization.
- Goldenbraid methodology: Batch cloning of synthesized promoter variants into plasmids.
- Protoplast transfection: Conducted in 96-well plates, enabling high throughput testing.
- Quantitative analysis: Luminescence measured using a microplate reader.
- Scalability: Capable of analyzing numerous promoter variants swiftly, with potential for stable transformation into plants for further validation.
Technology readiness level
This innovative assay platform is at Technology Readiness Level 5, indicating that the technology has been validated in relevant environments, specifically through high throughput testing in protoplasts. Future development involves collaboration with partners to design and synthesize promoter libraries for specific applications, with potential stable integration into plant genomes for further validation.
About University of North Texas
The University of North Texas is a comprehensive public research university in Denton, part of the UNT System, serving a large and diverse student body with broad academic programs. A dedicated research campus brings engineering and science together with shared user facilities, advanced instrumentation, prototyping spaces, and technology transfer support that speed collaboration with industry. Its Dallas–Fort Worth location offers ready access to major corporate R&D, suppliers, and testing partners, supported by project-based engagements, internships, and sponsored capstones. Research is backed by competitive federal funding from agencies such as the National Science Foundation, Department of Defense, and Department of Energy, alongside state and industry support.
Inquire about this partnering listing
Industry applications
delivered to your inbox.