Chapters authored
Rapid, High-Throughput Detection of Endocrine Disrupting Chemicals Using Autobioluminescent Cellular Bioreporters
Overexposure to endocrine disruptor chemicals (EDCs) can result in serious health problems, yet they are commonly found in everyday items such as pesticides, personal care products, nutritional supplements, and plastics. The U.S. Environmental Protection Agency, along with other such agencies from around the world, have therefore mandated that new approaches be designed to screen these products for the presence of EDCs. However, despite the presence of several types of extant EDC detection assays, there still exists a backlog approaching 87,000 chemicals currently awaiting screening. Autobioluminescent detection systems, which utilize cellular bioreporters capable of autonomously modulating bioluminescent signals without the need for external stimulation or investigator interaction, provide an attractive means for addressing this backlog because of their reduced performance costs and increased throughput relative to alternative assay systems. This chapter reviews the variety of existing EDC detection assays and evaluates the performance of a representative autobioluminescent estrogen-responsive EDC bioreporter to provide an overview of how autobioluminescence can be used to improve EDC detection using in vitro assay systems.
Part of the book: Endocrine Disruptors
Biotechnological Advances in Luciferase Enzymes
This chapter explores the history of the bioengineering advances that have been applied to common luciferase enzymes and the improvements that have been accomplished by this work. The primary focus is placed on firefly luciferase (FLuc), Gaussia luciferase (GLuc), Renilla luciferase (RLuc), Oplophorus luciferase (OLuc; NanoLuc), and bacterial luciferase (Lux). Beginning with the cloning and exogenous expression of each enzyme, their step-wise modifications are presented and the new capabilities endowed by each incremental advancement are highlighted. Using the historical basis of this information, the chapter concludes with a prospective on the overall impact these advances have had on scientific research and provides an outlook on what capabilities future advances could unlock.
Part of the book: Bioluminescence