Tools4Cells: Developing SCRIBE, a new programmable reversible transcriptional switch

The goal of this project is to build a new synthetic tool that will provide researchers with a greater degree of control over gene expression in plants. The major innovation of this tool is the ability to turn plant genes on and off in the desired location and in a highly controlled manner. The key is the ability to control tissue-specific expression of plant signals such as auxin, which will in turn lead to expression of the desired set of tissue-specific genes. This greater level of gene expression control will be important for the development of many advanced biotechnologies and has potential applications in agriculture such as the improvement of fruit or cereal yield. This project will facilitate the training of high school teachers and the next generation of scientists, and be at the center of K-12 community engagement programs. The development of molecular tools to control and monitor the transcriptional activity of a gene has had a tremendous impact not only on basic biological research but also on applied biotechnology and biomedicine. Two of the main drivers of recent technological advances in the area of transcriptional control have been the adoption of easily programmable DNA-binding proteins based on nuclease-inactivated Clustered Regularly Interspaced Short Palindromic Repeats systems and highly efficient transcriptional switches based on site-specific recombinases. Despite their utility, both of these technologies suffer from certain handicaps. To overcome this limitation, a new transcriptional control device is proposed which combines the flexibility, scalability, and programmability of the Clustered Regularly Interspaced Short Palindromic Repeats systems with the on/off and memory capabilities of the recombinase-based frameworks. The unique properties of this tool will make currently challenging studies, such as the precise spatiotemporal recording of transcriptional events in a whole organism, much more straightforward and informative. Importantly, the modularity of the system will allow for the facile customization of the device, tailoring it to specific experimental needs by simply "plugging" different input and output modules into the core switch. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria. NSF Award ID: 2533334 | Program: 01002627DB NSF RESEARCH & RELATED ACTIVIT | Principal Investigator: Jose Alonso | Institution: North Carolina State University, RALEIGH, NC | Award Amount: $853,932 View on NSF Award Search: https://www.nsf.gov/awardsearch/show-award/?AWD_ID=2533334 View on Research.gov: https://www.research.gov/awardapi-service/v1/awards/2533334.html