Decreased Levels of the RNA Splicing Factor Prpf3 in Mice and Zebrafish Do Not Cause Photoreceptor Degeneration
- Graziotto, J.J., Inglehearn, C.F., Pack, M.A., and Pierce, E.A.
- Investigative ophthalmology & visual science 49(9): 3830-3838 (Journal)
- Registered Authors
- Pack, Michael
- MeSH Terms
- Cell Line
- DNA Primers
- Embryonic Stem Cells/physiology*
- RNA Precursors/metabolism*
- Retinal Cone Photoreceptor Cells/pathology*
- Retinal Rod Photoreceptor Cells/pathology*
- Ribonucleoprotein, U4-U6 Small Nuclear/deficiency
- Ribonucleoprotein, U4-U6 Small Nuclear/metabolism*
- 18552388 Full text @ Invest. Ophthalmol. Vis. Sci.
Graziotto, J.J., Inglehearn, C.F., Pack, M.A., and Pierce, E.A. (2008) Decreased Levels of the RNA Splicing Factor Prpf3 in Mice and Zebrafish Do Not Cause Photoreceptor Degeneration. Investigative ophthalmology & visual science. 49(9):3830-3838.
Purpose: Pre-mRNA Processing Factor 3 (PRPF3) is a spliceosomal component essential for pre-mRNA processing. Mutations in PRPF3 have been implicated in retinitis pigmentosa 18 (RP18) through an unknown mechanism. We created and characterized Prpf3 knockout mice and zebrafish to determine if RP18 is a result of haploinsufficiency. Methods: Mice were produced from a Prpf3 genetrap cell line, and parameters of retinal function, structure, and RNA splicing were analyzed. The retinas of prpf3 insertional mutant zebrafish were also analyzed histologically. Results: Homozygous Prpf3 knockout mice do not survive to 14dpf, implying that this allele is required for early embryonic development. Homozygous Prpf3 knockout zebrafish die by 4dpf, well beyond the mid-blastula transition at which transcription activates. Zebrafish knockout embryos reveal abnormally high levels of cell death in the developing eye. Heterozygous Prpf3 knockout mice have less than the expected 50% reduction in Prpf3 at both the mRNA and protein levels, implying compensatory expression from the wild-type allele. The heterozygous mice develop normally, with no changes in retinal function, no evidence for photoreceptor degeneration at up to 23 months of age and no decrease in pre-mRNA splicing of transcripts mutated in other forms of RP in the retina. Similarly, heterozygous prpf3 knockout zebrafish develop normally and show no retinal degeneration at up to 12 months of age. Conclusions: These models suggest that RP18 is not a result of haploinsufficiency, but instead arises from a toxic gain of function due to missense mutations in PRPF3.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes