Functionally Complete Excision of Conditional Alleles in the Mouse Suprachiasmatic Nucleus by Vgat-ires-Cre
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...
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Veröffentlicht in: | Journal of biological rhythms 2018-04, Vol.33 (2), p.179-191 |
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Sprache: | eng |
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Zusammenfassung: | 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+;
Clockfl/fl; Npas2m/m mice and Vgat-Cre+; Bmal1fl/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. |
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ISSN: | 0748-7304 1552-4531 |
DOI: | 10.1177/0748730418757006 |