Abrupt onset of mutations in a developmentally regulated gene during terminal differentiation of post-mitotic photoreceptor neurons in mice

Ivette M. Sandoval, Brandee A. Price, Alecia K. Gross, Fung Chan, Joshua D. Sammons, John H. Wilson, Theodore G. Wensel

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

For sensitive detection of rare gene repair events in terminally differentiated photoreceptors, we generated a knockin mouse model by replacing one mouse rhodopsin allele with a form of the human rhodopsin gene that causes a severe, early-onset form of retinitis pigmentosa. The human gene contains a premature stop codon at position 344 (Q344X), cDNA encoding the enhanced green fluorescent protein (EGFP) at its 3′ end, and a modified 5′ untranslated region to reduce translation rate so that the mutant protein does not induce retinal degeneration. Mutations that eliminate the stop codon express a human rhodopsin-EGFP fusion protein (hRho-GFP), which can be readily detected by fluorescence microscopy. Spontaneous mutations were observed at a frequency of about one per retina; in every case, they gave rise to single fluorescent rod cells, indicating that each mutation occurred during or after the last mitotic division. Additionally, the number of fluorescent rods did not increase with age, suggesting that the rhodopsin gene in mature rod cells is less sensitive to mutation than it is in developing rods. Thus, there is a brief developmental window, coinciding with the transcriptional activation of the rhodopsin locus, in which somatic mutations of the rhodopsin gene abruptly begin to appear.

Original languageEnglish (US)
Article numbere108135
JournalPloS one
Volume9
Issue number9
DOIs
StatePublished - Sep 29 2014

Fingerprint

rhodopsin
Rhodopsin
photoreceptors
Neurons
rods (retina)
Genes
neurons
mutation
Mutation
mice
genes
stop codon
green fluorescent protein
somatic mutation
Retinal Degeneration
Retinitis Pigmentosa
Terminator Codon
Nonsense Codon
5' Untranslated Regions
Fluorescence microscopy

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Sandoval, I. M., Price, B. A., Gross, A. K., Chan, F., Sammons, J. D., Wilson, J. H., & Wensel, T. G. (2014). Abrupt onset of mutations in a developmentally regulated gene during terminal differentiation of post-mitotic photoreceptor neurons in mice. PloS one, 9(9), [e108135]. https://doi.org/10.1371/journal.pone.0108135

Abrupt onset of mutations in a developmentally regulated gene during terminal differentiation of post-mitotic photoreceptor neurons in mice. / Sandoval, Ivette M.; Price, Brandee A.; Gross, Alecia K.; Chan, Fung; Sammons, Joshua D.; Wilson, John H.; Wensel, Theodore G.

In: PloS one, Vol. 9, No. 9, e108135, 29.09.2014.

Research output: Contribution to journalArticle

Sandoval, Ivette M. ; Price, Brandee A. ; Gross, Alecia K. ; Chan, Fung ; Sammons, Joshua D. ; Wilson, John H. ; Wensel, Theodore G. / Abrupt onset of mutations in a developmentally regulated gene during terminal differentiation of post-mitotic photoreceptor neurons in mice. In: PloS one. 2014 ; Vol. 9, No. 9.
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