Supplementary Figure 5: A20mZnF7/mZnF7 knock-in mice generated using CRISPR/Cas9-mediated gene targeting develop dactylitis. | Nature Cell Biology

Supplementary Figure 5: A20mZnF7/mZnF7 knock-in mice generated using CRISPR/Cas9-mediated gene targeting develop dactylitis.

From: A20 prevents inflammasome-dependent arthritis by inhibiting macrophage necroptosis through its ZnF7 ubiquitin-binding domain

Supplementary Figure 5

(a) Schematic representation of the A20 gene indicating the location of the point mutations introduced in exon 9 to change cysteine (C) residues at positions 764 and 767 to alanines (A). Two silent mutations were also introduced to avoid re-editing of the DNA by Cas9 after homology-directed repair. Sanger sequencing of wt and A20mZnF7/mZnF7 mouse tail DNA to confirm correct introduction of the designed mutations into the genome. (b) mCT analysis of forepaws of mice with the indicated genotypes depicts the extensive bone erosions in the digits of A20mZnF7/mZnF7 and the milder erosions in carpal bones compared to A20MYC-KO mice (c) Representative histological images from forepaws (right panel; transversal sections) and hindpaws (left panel; sagittal sections) of mice with the indicated genotypes. Note the development of dactylitis in A20mZnF7/mZnF7 mice with a characteristic severe tenosynovitis leading to disruption of muscle/tendon fibres and the pannus orchestrating destruction of bones (arrow), all being more evident in the distal and intermediate (arrowhead) than in proximal phalanxes (scale bar: 1mm) (A20wt/mZnF7, n=8 and A20mZnF7/mZnF7, n=10 mice).

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