Receptor knock-out and knock-in strategies

Methods in Molecular Biology
Ikuo Matsuda, Atsu Aiba

Abstract

Accumulating examples have demonstrated that knock-out and knock-in mice of G-protein-coupled receptors (GPCRs) are useful in elucidating physiological functions of the receptor in vivo. GPCR knock-out and knock-in are achieved by either (1) manipulation of the endogenous locus of the receptor gene or (2) transgenic expression of the modified receptor. Historically speaking, the first generation knock-outs made the best use of homologous recombination in embryonic stem (ES) cells and their totipotency to introduce the desired mutation into the endogenous receptor locus. In the second-generation knock-outs using the Cre/loxP system, the disruption of the receptor gene is cell-type specific or region-specific but is irreversible in principle. In contrast, transgenic expression in the receptor knock-out mice of the wild-type receptor protein under a tissue- and stage-specific promoter (conditional "rescue" of the receptor knock-out) can be easily applied to create "reversible" or "inducible" knock-out of the receptor. This is called the third generation knock-out. In the following sections, we introduce examples of the materials and methods based on our in vivo analyses of the metabotropic glutamate receptor-subtype 1 (mGluR1).

Citations

May 5, 2011·World Journal of Biological Chemistry·Helena HuangRuth M Empson

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