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.
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Acknowledgments
I am grateful to P. Mombaerts for critical reading the manuscript, M. Omura for photos of ES cells and valuable comments, S. Nishimura for comments on chromosome engineering, and members of the Mombaerts laboratory for improvements in the methods. This work was supported in part by JSPS KAKENHI Grant Number 24500456.
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Ishii, T. (2013). Genetic Manipulation to Analyze Pheromone Responses: Knockouts of Multiple Receptor Genes. In: Touhara, K. (eds) Pheromone Signaling. Methods in Molecular Biology, vol 1068. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-619-1_10
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DOI: https://doi.org/10.1007/978-1-62703-619-1_10
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