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Methods Enzymol

Title:		Silencing of OBP genes: Generation of loss-of-function mutants of PBP by genome editing
Author(s):		Shiota Y; Sakurai T;
Address:		"Department of Evolutionary Neuroethology, Max Planck Institute for Chemical Ecology, Jena, Germany; Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan. Faculty of Agriculture, Tokyo University of Agriculture, Kanagawa, Japan. Electronic address: ts206448@nodai.ac.jp"
Journal Title:		Methods Enzymol
Year:		2020
Volume:		20200627
Issue:
Page Number:		325 - 344
DOI:		10.1016/bs.mie.2020.05.009
ISSN/ISBN:		1557-7988 (Electronic) 0076-6879 (Linking)
Abstract:		"Pheromone binding proteins (PBPs) are small soluble proteins (about 15kDa) that play striking roles in the detection of sex pheromones in insects. Many studies including structural analysis, binding simulation, and in vitro assays have been performed to clarify the modes of action of PBPs. Although these studies have provided valuable contributions toward the understanding of which key amino acid components contribute to the correct folding of PBPs and their binding affinities to sex pheromones, the functional characteristics of PBPs in the natural environment is still obscure. Recent developments in genome editing have begun to enable the functional examination of PBPs in in vivo. Among insect PBPs, BmPBP1 is one of the most well-characterized, there being rich understanding of its structure, biochemical analysis, binding affinity, localization, and the relationship between the type of olfactory receptors and its expression. A recent study has shown that BmPBP1 contributes sensitivity, but not selectivity of sex pheromone detection in the silkmoth Bombyx mori. In this chapter, based on a current report of the functional characterization of BmPBP1 using genome editing, we provide one example of a useful analytical method to clarify the functional role of PBP in vivo"
Keywords:		"Animals *Bombyx/genetics Gene Editing Insect Proteins/genetics *Receptors, Odorant/genetics *Sex Attractants Bombyx mori Electroantennogram Functional characterization Genome editing Pheromone Pheromone binding protein Transcription activator-like effecto;"
Notes:		"MedlineShiota, Yusuke Sakurai, Takeshi eng Research Support, Non-U.S. Gov't 2020/08/24 Methods Enzymol. 2020; 642:325-344. doi: 10.1016/bs.mie.2020.05.009. Epub 2020 Jun 27"