Phenotype Image Detail

Image

Caption

Figure 1. Generation of a mouse model of CENPJ-Seckel syndrome. A. The CENPJ gene spans 40 kb and comprises 17 exons. The 3' and 5' untranslated regions are depicted in grey. Mutations in CENPJ have been associated with either primary microcephaly (MCPH) or Seckel syndrome (SECKEL). The mutation in intron 11 that has been associated with Seckel syndrome results in the generation of three transcripts: one lacking exon 12, one lacking 11 and 12 and one lacking exons 11,12 and 13. Disruption of mouse Cenpj by insertion of a cassette (depicted by the blue square) between exons 4 and 5 results in low levels of splicing over the cassette and cryptic splicing between exons 3 and 6 or 4 and 6; the latter two transcripts are predicted to result in truncated proteins. The allele was designated Cenpjtm1a(EUCOMM)Wtsi and abbreviated to Cenpjtm. Percentages show mean expression of Cenpj across exon boundaries as determined by quantitative RT-PCR relative to Gapdh for Cenpj^tm/tm relative to Cenpj^+/+ for RNA extracted from n = 3 murine embryonic fibroblast (MEF) lines. B. Immunoblot to show reduction in Cenpj levels in protein extracted from Cenpj^tm/tm (^tm/tm), Cenpj^+/tm (^+/tm), and Cenpj^+/+(^+/+) MEFs. KAP1 was used as a loading control. C. Table shows frequency of Cenpj^tm/tm mice born from heterozygote intercrosses. Cenpj^tm/tm showed partial embryonic lethality as shown by their reduced frequency at E18.5 and P14 (25% expected, *P = 0.02, **P = 0.0001, chi squared test). D. Representative images of E18.5 skeletal preparations of Cenpj^+/+ and Cenpj^tm/tm embryos. Staining with alcian blue (cartilage) and alizarin red (bone). Cenpj^tm/tm embryo with a sloping forehead and polysyndactylism of digit one of the left hindpaw (inset). E. Bodyweights of male Cenpj^tm/tm (n = 8), Cenpj^+/tm (n = 7), Cenpj^+/+(n = 40) and baseline wild-type controls (n = 912) from 3-16 weeks of age. Data show that Cenpj^tm/tm are significantly smaller than Cenpj^+/+ mice at all ages (P = 2.2x10-16, Mann-Whitney-Wilcoxon test). F. Skeletal preparations of E18.5 Cenpj^tm/tm embryos showed irregular ossification of the cranium and G. sternum. H. X-Rays show that adult Cenpj^tm/tm mice may present with a flatter, sloping forehead (A), mild elevation of the parietal bone (B), a short humerus with a prominent deltoid tuberosity (C), prominent medial epicondyle (D), an irregular ribcage (E), short lumbar and sacral vertebrae (F), an abnormal pelvis (G), extra sacrocaudal transitional vertebrae (H), short, abnormal/fused caudal vertebrae 2/3 - caudal vertebrae 7/8 (I) and reduced intervertebral joint space (J).

Copyright

This image is from McIntyre RE, PLoS Genet 2012;8(11):e1003022, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:194085

Associated Alleles

Symbol	Name
Cenpj^tm1a(EUCOMM)Wtsi	centromere protein J; targeted mutation 1a, Wellcome Trust Sanger Institute
Cenpj^tm1a(EUCOMM)Wtsi	centromere protein J; targeted mutation 1a, Wellcome Trust Sanger Institute

Associated Genotypes

Allelic Composition	Genetic Background
Cenpj^tm1a(EUCOMM)Wtsi/Cenpj^tm1a(EUCOMM)Wtsi	B6Brd;B6N-Tyr^c-Brd Cenpj^tm1a(EUCOMM)Wtsi/Wtsi