clock homolog (mouse) (CLOCK),

Manipulation of Mammalian Cell Lines for Circadian Studies

and culture a fibroblast cell line from wild-type and knockout mouse skin and (2) transfect cells at high efficiency to use in functional clock-protein studies.

Microinjection of In Vitro Transcribed RNA and Antisense Oligonucleotides in Mouse Oocytes and Early Embryos to Study the Gain-and Loss-of-Function of

injected mRNAs with high fidelity. However, Fig. 1 . Microinjection of HSP70 antisense mRNA into one-cell mouse embryos results in inhibition of HSP7O expression. Immunoprecipitation was carried out pith an HSP70 antibody and the immunoprecipitate resolved

Coimmunoprecipitation Assay

As with most other proteins, clock proteins physically interact with one another. Coimmunoprecipitation (coIP) is the most straightforward technique to study protein-protein interactions in vivo, if antibodies against the proteins