CRISPR/Cas9‐mediated deletion of Fam83h induces defective tooth mineralization and hair development in rabbits

Family with sequence similarity 83 members H (Fam83h) is essential for dental enamel formation. Fam83h mutations cause human amelogenesis imperfecta (AI), an inherited disorder characterized by severe hardness defects in dental enamel. Nevertheless, previous studies showed no enamel defects in Fam83h‐knockout/lacZ‐knockin mice. In this study, a large deletion of the Fam83h gene (900 bp) was generated via a dual sgRNA‐directed CRISPR/Cas9 system in rabbits. Abnormal tooth mineralization and loose dentine were found in homozygous Fam83h knockout (Fam83h−/−) rabbits compared with WT rabbits. In addition, reduced hair follicle counts in dorsal skin, hair cycling dysfunction and hair shaft differentiation deficiency were observed in Fam83h−/− rabbits. Moreover, X‐rays and staining of bone sections showed abnormal bending of the ulna and radius and an ulnar articular surface with insufficient trabecular bone in Fam83h−/− rabbits. Taken together, these data are the first report of defective hair cycling, hair shaft differentiation and abnormal bending of the ulna and radius in Fam83h−/− rabbits. This novel Fam83h−/− rabbit model may facilitate understanding the function of Fam83h and the pathogenic mechanism of the Fam83h mutation.

[1]  M. Wolf,et al.  FGF23 at the crossroads of phosphate, iron economy and erythropoiesis , 2019, Nature Reviews Nephrology.

[2]  T. Porntaveetus,et al.  Decreased osteogenic activity and mineralization of alveolar bone cells from a patient with amelogenesis imperfecta and FAM83H 1261G>T mutation , 2019, Genes & diseases.

[3]  B. Baban,et al.  Generation of Fam83h knockout mice by CRISPR/Cas9‐mediated gene engineering , 2019, Journal of cellular biochemistry.

[4]  Jung‐Wook Kim,et al.  The Enamel Phenotype in Homozygous Fam83h Truncation Mice , 2019, Molecular genetics & genomic medicine.

[5]  L. Lai,et al.  The disrupted balance between hair follicles and sebaceous glands in Hoxc13‐ablated rabbits , 2018, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[6]  M. Fan,et al.  DLX3 Inhibits the Proliferation of Human Dental Pulp Cells Through Inactivation of Canonical Wnt/β-Catenin Signaling Pathway , 2018, Front. Physiol..

[7]  Yang Liu,et al.  A novel FAM83H mutation in one Chinese family with autosomal-dominant hypocalcification amelogenesis imperfecta , 2018, Mutagenesis.

[8]  G. Sapkota,et al.  The FAM83 family of proteins: from pseudo-PLDs to anchors for CK1 isoforms , 2018, Biochemical Society transactions.

[9]  Tingting Zhang,et al.  Fam83h mutation inhibits the mineralization in ameloblasts by activating Wnt/β-catenin signaling pathway. , 2018, Biochemical and biophysical research communications.

[10]  G. Sapkota,et al.  The DUF1669 domain of FAM83 family proteins anchor casein kinase 1 isoforms , 2018, Science Signaling.

[11]  C. Sorenson,et al.  Mice dental pulp and periodontal ligament endothelial cells exhibit different proangiogenic properties. , 2018, Tissue & cell.

[12]  G. Yuan,et al.  BMP-2 induced Dspp transcription is mediated by Dlx3/Osx signaling pathway in odontoblasts , 2017, Scientific Reports.

[13]  C. Inglehearn,et al.  Amelogenesis Imperfecta; Genes, Proteins, and Pathways , 2017, Front. Physiol..

[14]  R. Boyd,et al.  A novel Foxn1eGFP/+ mouse model identifies Bmp4‐induced maintenance of Foxn1 expression and thymic epithelial progenitor populations , 2017, European journal of immunology.

[15]  T. Tomonaga,et al.  FAM83H and casein kinase I regulate the organization of the keratin cytoskeleton and formation of desmosomes , 2016, Scientific Reports.

[16]  L. Lai,et al.  CRISPR/Cas9-mediated mutation of PHEX in rabbit recapitulates human X-linked hypophosphatemia (XLH). , 2016, Human molecular genetics.

[17]  Murim Choi,et al.  Fam83h null mice support a neomorphic mechanism for human ADHCAI , 2015, Molecular genetics & genomic medicine.

[18]  Wei Zhang,et al.  Hoxc13 is a crucial regulator of murine hair cycle , 2015, Cell and Tissue Research.

[19]  Misako Sato,et al.  A novel mechanism of keratin cytoskeleton organization through casein kinase I&agr; and FAM83H in colorectal cancer , 2013, Journal of Cell Science.

[20]  Le Cong,et al.  Multiplex Genome Engineering Using CRISPR/Cas Systems , 2013, Science.

[21]  Jeffrey C. Miller,et al.  A rapid and general assay for monitoring endogenous gene modification. , 2010, Methods in molecular biology.

[22]  C. Giachelli,et al.  The role of osteopontin in inflammatory processes , 2009, Journal of Cell Communication and Signaling.

[23]  T. Hart,et al.  Phenotypic Variation in FAM83H-associated Amelogenesis Imperfecta , 2009, Journal of dental research.

[24]  Jung‐Wook Kim,et al.  Mutational spectrum of FAM83H: the C‐terminal portion is required for tooth enamel calcification , 2008, Human mutation.

[25]  F. Giráldez,et al.  BMP-signaling regulates the generation of hair-cells. , 2006, Developmental biology.

[26]  G. Stein,et al.  Dlx3 Transcriptional Regulation of Osteoblast Differentiation: Temporal Recruitment of Msx2, Dlx3, and Dlx5 Homeodomain Proteins to Chromatin of the Osteocalcin Gene , 2004, Molecular and Cellular Biology.

[27]  C. Chuong,et al.  `Cyclic alopecia' in Msx2 mutants: defects in hair cycling and hair shaft differentiation , 2003, Development.

[28]  M. Shen,et al.  Msx homeobox genes inhibit differentiation through upregulation of cyclin D1. , 2001, Development.

[29]  B. Hogan,et al.  Inhibition of Bmp signaling affects growth and differentiation in the anagen hair follicle , 2000, The EMBO journal.

[30]  C. D. Cox,et al.  Effect of red blood cells on the growth of Porphyromonas endodontalis and microbial community development. , 1998, Oral microbiology and immunology.