Functionally Complete Excision of Conditional Alleles in the Mouse Suprachiasmatic Nucleus by Vgat-ires-Cre
Department of Neurobiology; Program in Neuroscience, Graduate School of Biomedical Sciences; Weaver Lab; Anaclet Lab
Neuroscience and Neurobiology
Mice with targeted gene disruption have provided important information about the molecular mechanisms of circadian clock function. A full understanding of the roles of circadian-relevant genes requires manipulation of their expression in a tissue-specific manner, ideally including manipulation with high efficiency within the suprachiasmatic nuclei (SCN). To date, conditional manipulation of genes within the SCN has been difficult. In a previously developed mouse line, Cre recombinase was inserted into the vesicular GABA transporter (Vgat) locus. Since virtually all SCN neurons are GABAergic, this Vgat-Cre line seemed likely to have high efficiency at disrupting conditional alleles in SCN. To test this premise, the efficacy of Vgat-Cre in excising conditional (fl, for flanked by LoxP) alleles in the SCN was examined. Vgat-Cre-mediated excision of conditional alleles of Clock or Bmal1 led to loss of immunostaining for products of the targeted genes in the SCN. Vgat-Cre(+); Clock(fl/fl); Npas2(m/m) mice and Vgat-Cre(+); Bmal1(fl/fl) mice became arrhythmic immediately upon exposure to constant darkness, as expected based on the phenotype of mice in which these genes are disrupted throughout the body. The phenotype of mice with other combinations of Vgat-Cre(+), conditional Clock, and mutant Npas2 alleles also resembled the corresponding whole-body knockout mice. These data indicate that the Vgat-Cre line is useful for Cre-mediated recombination within the SCN, making it useful for Cre-enabled technologies including gene disruption, gene replacement, and opto- and chemogenetic manipulation of the SCN circadian clock.
circadian rhythms, Cre recombinase, conditional gene disruption, Clock, Npas2, Bmal1, Scl32a1, GABAergic neurons
DOI of Published Version
J Biol Rhythms. 2018 Apr;33(2):179-191. doi: 10.1177/0748730418757006. Link to article on publisher's site
Journal of biological rhythms
Weaver, David R.; van der Vinne, Vincent; Giannaris, Eustathia Lela; Vajtay, Thomas J.; Holloway, Kristopher L.; and Anaclet, Christelle, "Functionally Complete Excision of Conditional Alleles in the Mouse Suprachiasmatic Nucleus by Vgat-ires-Cre" (2018). Neurobiology Publications and Presentations. 223.