Red-Shifted FRET Biosensors for High-Throughput Fluorescence Lifetime Screening

We have developed fluorescence resonance energy transfer (FRET) biosensors with red-shifted fluorescent proteins (FP), yielding improved characteristics for time-resolved (lifetime) fluorescence measurements. In comparison to biosensors with green and red FRET pairs (GFP/RFP), FPs that emit at longer wavelengths (orange and maroon, OFP/MFP) increased the FRET efficiency, dynamic range, and signal-to-background of high-throughput screening (HTS). OFP and MFP were fused to specific sites on the human cardiac calcium pump (SERCA2a) for detection of structural changes due to small-molecule effectors. When coupled with a recently improved HTS fluorescence lifetime microplate reader, this red-shifted FRET biosensor enabled high-precision nanosecond-resolved fluorescence decay measurements from microliter sample volumes at three minute read times per 1536-well-plate. Pilot screens with a library of small-molecules demonstrate that the OFP/MFP FRET sensor substantially improves HTS assay quality. These high-content FRET methods detect minute FRET changes with high precision, as needed to elucidate novel structural mechanisms from small-molecule or peptide regulators discovered through our ongoing HTS efforts. FRET sensors that emit at longer wavelengths are highly attractive to the FRET biosensor community for drug discovery and structural interrogation of new therapeutic targets

Measuring Innovation in Carver County

Report completed by students enrolled in MBA 6220: Operations Management, taught by Scott Martens in spring 2016.This project was completed as part of the 2015-2016 Resilient Communities Project (rcp.umn.edu) partnership with Carver County. Carver County staff had initiated continuous improvement and innovation workshops across a variety of departments. The goal of this project was to identify evaluation tools to assess the impact of these various continuous improvement efforts. Carver County project lead Lorraine Brady worked with nine teams of students in MBA 6220: Operations and Management to evaluate the initiatives, identify measurement tools and strategies, and develop recommendations for continuing improvement actions implemented during the workshops. The students' aggregated final report is available.This project was supported by the Resilient Communities Project (RCP), a program at the University of Minnesota whose mission is to connect communities in Minnesota with U of MN faculty and students to advance local sustainability and resilience through collaborative, course-based projects. RCP is a program of the Center for Urban and Regional Affairs (CURA). More information at http://www.rcp.umn.edu.Martens, Scott; Bawaskar, Prachi; Liang, Jack; Sawatzky, Jared; Wagle, David; Bell, Carly; Collins, Lindsey; Grant, Dan; Kinsella, Mike; Lonergan, Brendan; Mueller, Michaela; Schober, Marc; Blake, Katie; Daugherty, Molly; Halsch, Ryan; Kitchen, Ian; Lund, Jon; Shah, Rahul; White, Liz; Zimmermann, Katherine; Berghoff, Jeremy; Drott, D. Stephen; Haun, Trent; Lundstrom, Joel; Preston, Sean; Bowersock, Alyssa; Dunlap, Claire; Hektner, Michael; Korman, Daniel; Lusk, Jason; Reveland, Nick; Sohn, Garrett; Butskiy, Konstantin; Dyshaw, Brittany; Hinton, Sam; Kler, Param; Novosad, Alana; Saladi, Siddhartha; Tawfik, Daniel; El-Sawaf, Khaled; Hulke, Mandy; Lamuro, Margaret; Sansone, Rebecca; Byers, Russell; McCormick, Timothy; Daugherty, Laura; Hendrickson, Briana; Kyanam, Devi; Larson, Sam; Madetzke, Mike; Mcelhinny, Ryan; Saprygina, Polina; Thai, Thuong. (2016). Measuring Innovation in Carver County. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/184940