Conditional allelic replacement applied to genes encoding the histone variant H3.3 in the mouse.

Michelle C W Tang, Shelley A Jacobs, Lee H Wong, Jeffrey R Mann

Theme of Genetic Disorders, Murdoch Childrens Research Institute, The Royal Children's Hospital, Victoria, 3052, Australia.

Genesis (New York, N.Y. : 2000) 2013 Feb

filter terms:

Post-translational modifications to residues in core histones convey epigenetic information. Their function can be evaluated in amino acid substitution mutants, although to date this method has not been used in mice. To this end, we have evaluated gene targeting vectors designed for Cre recombinase-mediated conditional allelic replacement at the two unlinked genes encoding the histone variant H3.3. The conditional alleles consist of an uninterrupted wild-type H3.3 coding sequence upstream of a desired alternative or proxy coding sequence. The arrangement of two loxP sites allows Cre-mediated replacement of the wild-type coding sequence with the proxy. To demonstrate proof of principle, at each locus we replaced the wild-type coding sequence with a fluorescent reporter. This produced null alleles that will be useful to analyse the effects of H3.3 deficiency in development. Each targeting vector can readily be retrofitted with a proxy coding sequence encoding a modified H3.3 protein. Such vectors will allow for the conditional substitution of specific residues in order to dissect the roles of H3.3 post-translational modifications in development and disease. Copyright © 2012 Wiley Periodicals, Inc.

Citation

Michelle C W Tang, Shelley A Jacobs, Lee H Wong, Jeffrey R Mann. Conditional allelic replacement applied to genes encoding the histone variant H3.3 in the mouse. Genesis (New York, N.Y. : 2000). 2013 Feb;51(2):142-6