In-Fusion® Cloning—a Fast, Efficient Way to Create Hundreds of Genetically-Encoded Biosensor Prototypes for Screening in Living Cells

2 Aug 2015

Genetically-encoded, fluorescent biosensors have made it possible to look at signaling events in living cells. The greatest difficulty in creating FRET-based biosensors is finding the right way to fuse the donor and acceptor fluorophores to signaling proteins, or protein domains, without disrupting protein or cellular function. This application note describes the In-Fusion strategy, making it possible for a small team to create hundreds of biosensor prototypes quickly and efficiently.

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BiosensorsBiosensors are devices used to detect an analyte using biological molecules specific to the analyte coupled to a detector. Biosensor instruments may be photometric, typically using surface plasma resonance (SPR), electrochemical or QCM (quartz crystal microbalance) biosensors. Biosensors should be selective, portable, robust and sensitive and have a fast response time. Systems may be manual or automated and usually have associated software.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein InteractionsProtein interactions are essential for understanding cellular processes, as proteins work together to carry out biological functions. Studying protein interactions can provide insights into disease mechanisms, drug development, and cell signaling pathways. Explore the best tools for protein interaction research in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Live Cell ImagingLive cell imaging is the study of living cells using microscopes and high-content imaging systems. This technique provides in-depth insight into fast and complex biological processes, by allowing dynamic imaging of living cells instead of acquiring an individual image at a single point in time.Biomarker DiscoveryClinical biomarkers refer to substances related to known medical conditions that can be accurately measured <i>in vitro</i>. Biomarkers can be used to diagnose presence of a disease and indicate disease severity. The discovery of new biomarkers is incredibly valuable in the field of diagnostics.Genetic EngineeringCell SignalingFRET