Fig. 1

Fig. 1 Patient pedigrees and genomic comparison of zf and human FAM222B. (A) Pedigrees of cases with cardiovascular laterality defects, situs inversus totalis, heterotaxy, or isolated CHD identified through exome sequencing. Circle for female, square for male. Doubled line for consanguineous marriage. Affected patients in black: I1: lifelong reduced physical capacity, late diagnosis of an unspecified ASD; II2: PDA, patent foramen ovale (PFO); II3: congenital valvular pulmonary stenosis; III1: VSD, PDA, ASD II, bicuspid aortic valve with stenosis and insufficiency; III2: multiple VSDs, double chambered right ventricle, PDA, bicuspid aortic valve with insufficiency; III3: VSD, PDA, PFO; III4: common truncus arteriosus type A1, ASD II, left superior vena cava. * = No DNA available (B) Variant table of FAM222B including specific changes in base and amino acid sequence, MAF, CADD and ACMG, VUS = variant of uncertain significance, * cannot be classified according to ACMG criteria. (C) Genetic structure of human FAM222B, the zf homologues fam222ba/bb and zf fam222aa. The introns (lines), exons (boxes) and protein coding (dark blue) sequence sizes were adjusted according to the entire gene. Dark green bands underneath the human FAM222B reveal the genetic conservation score compared to zf (UCSC genome browser). The six variants identified in our studies are marked with an arrow (point-mutations) or line (deletion) in the highly conserved exon three. Variant 1/2, which was further observed in our studies, was highlighted in red; other variants were highlighted in orange. Red bars in the zf paralogues reveal the position of the 10 bp deletion in the TALEN-generated fam222ba/bb dd-KO line, leading to a reading frame shift with a resulting early stop codon. Confirmative sequencing of the received dd-KO zfl is shown in the box aside. Blue numbers beneath each chromosome label represent a comparative analysis between human FAM222B and zf homologues Fam222ba, Fam222bb as well as zf Fam222aa on amino acid level using the UniProt align tool. The highest similarity was revealed between FAM222B and Fam222bb (59.9%), followed by Fam222ba (54.2%) and Fam222aa (34.6%).