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https://www.selleckchem.com/products/pf-07265807.html Cellular functions are controlled by sophisticated signal transduction pathways triggered by receptors responding to myriad environmental stimuli. With the rise of synthetic biology, we can now engineer artificial receptors enabling real-time interrogation and manipulation of cellular signaling, and providing new clues about the design principles of natural sensing systems. In this review, we describe the main classes of synthetic receptors engineered to date, their applications, and highlight recent developments that might improve synthetic receptor design in the future. © 2020 Elsevier Inc. All rights reserved.Nuclease-mediated DNA cleavage and subsequent repair lie at the heart of genome editing, and the RNA-guided endonuclease Cas9 has emerged as the most widely-used tool for facilitating this process. Extensive biochemical and biophysical efforts have revealed much regarding the structure, mechanism, and cellular properties of Cas9. This has enabled engineering of Cas9 variants with enhanced activity, specificity, and other features. However, we lack a detailed understanding of the kinetics of Cas9-mediated DNA cleavage and repair in vivo. To study in vivo Cas9 cleavage kinetics and activity dose-dependence, we have engineered a chemically-inducible, single-component Cas9, ciCas9. ciCas9 allows for temporal and rheostatic control of Cas9 activity using a small molecule activator, A115. We have also developed a droplet-digital PCR-based assay (DSB-ddPCR) to directly quantify Cas9-mediated double-stranded breaks (DSBs). The methods in this chapter describe the application of ciCas9 and DSB-ddPCR to study the kinetics and dose-dependence of Cas9 editing in vivo. © 2020 Elsevier Inc. All rights reserved.RAS GTPases are involved in a number of dynamic signaling processes and have been a major focus of research due to the prevalence of activating RAS mutations in cancer. However, despite decades of research, some

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