title: Genetic manipulation to analyze pheromone responses: knockouts of multiple receptor genes.
abstract: Gene targeting in the mouse is an essential technique to study gene function in vivo. Multigene families encoding vomeronasal receptor (VR) type 1 and type 2 consist of ~300 intact genes, which are clustered at multiple loci in the mouse genome. To understand the function of VRs and neurons expressing a particular VR in vivo, individual endogenous receptor genes can be manipulated by conventional gene targeting to create loss-of-function mutations or to visualize neurons and their axons expressing the VR. Multiple receptor genes in a cluster can also be deleted simultaneously by chromosome engineering, allowing analysis of function of a particular VR subfamily. Here, we describe protocols for conventional gene targeting and chromosome engineering for deleting a large genomic region in mouse embryonic stem (ES) cells.
authors.affiliation: Department of Cell Biology, Graduate School of Medical and Dental Science, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ishii
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology; D018390:Gene Targeting; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D005810:Multigene Family / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D043563:Receptors, Pheromone / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D019147:Vomeronasal Organ / Q000166:cytology / Q000378:metabolism*
chemical_list: D043562:Receptors, G-Protein-Coupled; D043563:Receptors, Pheromone