Titlebook: Notch Signaling Research; Methods and Protocol Dongyu Jia Book 2022 The Editor(s) (if applicable) and The Author(s), under exclusive licens

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1064-3745 expertsThis detailed volume explores the large research area of Notch signaling studies in a variety of model organisms. The collected research methods and protocols include the dissection of Notch functional sites, Notch regulators, Notch activity reporters and analysis, roles of Notch in developm

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Using the CRISPR/Cas9 System for Dissection of Functional Sites of the , Gene in , ,, which were replaced by the . integration site of the .C31 phage. Based on this approach, we obtained and characterized new . mutations. Thus, a new powerful tool is provided for studying the genetic regulation of the . gene and the organization of chromatin at this locus.

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Screening Mutants by Single Fly Genomic PCR,n all independent and individual excised mutant fly lines before the presence of a mutation is confirmed. Here, we provide a method to detect and confirm the presence of a mutation, as and when mutant flies are generated. By allowing for the maintenance and expansion of only the confirmed mutant lin

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Generation of Properly Folded Epidermal Growth Factor-Like (EGF) Repeats and Glycosyltransferases Ered proteins, including the Notch receptors. Notch receptors include numerous EGF repeats tandemly connected in the extracellular domain. These EGF repeats must be properly folded in order for them to undergo the three different types of .-glycosylation associated with these extracellular proteins:

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Artificial Notch Signaling Activation Method Using Immobilized Ligand Beads,cellular domain. Therefore, the soluble recombinant ligand protein is not suitable for the activation of Notch signaling in a cell culture system. Here, we describe an efficient method for transient activation of Notch signaling using immobilized ligand beads. Using this method, the timing of Notch

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