title: Relationship between somatic mosaicism of Pax6 mutation and variable developmental eye abnormalities-an analysis of CRISPR genome-edited mouse embryos.
abstract: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system is a rapid gene-targeting technology that does not require embryonic stem cells. To demonstrate dosage effects of the Pax6 gene on eye formation, we generated Pax6-deficient mice with the CRISPR/Cas system. Eyes of founder embryos at embryonic day (E) 16.5 were examined and categorized according to macroscopic phenotype as class 1 (small eye with distinct pigmentation), class 2 (pigmentation without eye globes), or class 3 (no pigmentation and no eyes). Histologically, class 1 eyes were abnormally small in size with lens still attached to the cornea at E16.5. Class 2 eyes had no lens and distorted convoluted retinas. Class 3 eyes had only rudimentary optic vesicle-like tissues or histological anophthalmia. Genotyping of neck tissue cells from the founder embryos revealed somatic mosaicism and allelic complexity for Pax6. Relationships between eye phenotype and genotype were developed. The present results demonstrated that development of the lens from the surface ectoderm requires a higher gene dose of Pax6 than development of the retina from the optic vesicle. We further anticipate that mice with somatic mosaicism in a targeted gene generated by CRISPR/Cas-mediated genome editing will give some insights for understanding the complexity in human congenital diseases that occur in mosaic form.
authors.affiliation: Department of Orthodontics Dentofacial Orthopedics, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8504, Japan. yasuea@tokushima-u.ac.jp.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yasue
authors.affiliation: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Kono
authors.affiliation: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan.
authors.forename: Munenori
authors.initials: M
authors.lastname: Habuta
authors.affiliation: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Bando
authors.affiliation: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan.
authors.forename: Keita
authors.initials: K
authors.lastname: Sato
authors.affiliation: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan.
authors.forename: Junji
authors.initials: J
authors.lastname: Inoue
authors.affiliation: Division of Molecular Biology, Institute for Advanced Enzyme Research, Tokushima University, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Oyadomari
authors.affiliation: Tokushima University, 2-24 Shinkura-cho, Tokushima, 770-8501, Japan.
authors.forename: Sumihare
authors.initials: S
authors.lastname: Noji
authors.affiliation: Department of Orthodontics Dentofacial Orthopedics, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8504, Japan.
authors.forename: Eiji
authors.identifier: 0000-0002-2707-1613
authors.initials: E
authors.lastname: Tanaka
authors.affiliation: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8558, Japan. hohuchi@okayama-u.ac.jp.
authors.forename: Hideyo
authors.initials: H
authors.lastname: Ohuchi
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D004475:Ectoderm; D004622:Embryo, Mammalian; D005136:Eye Proteins / Q000235:genetics*; D018628:Gene Dosage; D000072669:Gene Editing; D018398:Homeodomain Proteins; D007908:Lens, Crystalline / Q000002:abnormalities* / Q000196:embryology; D008822:Mice, Transgenic; D008850:Microphthalmos / Q000196:embryology / Q000235:genetics; D009030:Mosaicism; D000071841:PAX6 Transcription Factor / Q000235:genetics*; D010641:Phenotype; D015792:Retinal Dysplasia / Q000196:embryology / Q000235:genetics
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor