case_images: PMC10485611_01
case_images: Abnormal imaging of the proband. (A) X-ray of the proband showing a short chest and distorted and fused ribs. Chest CT demonstrating hemivertebrae (B, upper arrow), butterfly vertebrae (B, lower arrow), rib fusion (C), and spinal canal enlargement (D,E) Kidney color Doppler ultrasound showing left hydronephrosis. (F) Chest x-ray showing mild scoliosis at the age of 11 months.
case_images: fped-11-1201999-g001.jpg
case_images: PMC10485611_01_fped-11-1201999-g001.jpg
case_images: F1
case_images: Mild scoliosis was observed on the chest x-ray (Figure 1F). The kinship value between the couple was 0.0721 which indicated that they were third degree relatives (Supplementary Figure S1).
case_images: Genetic analysis of the HES7 gene. (A) Pedigree diagram and (B) Sanger sequencing results of the family. (C) Transcripts of the HES7 gene. (D) Overview of the reported HES7 variants.
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case_images: PMC10485611_01_fped-11-1201999-g002.jpg
case_images: F2
case_images: A homozygous variant HES7 c.226+1G>A was identified by trio-WES, which was inherited from both parents (Figures 2A,B). HES7 is located at chromosome 17p13.1 and has two transcripts, namely, NM_001165967 and NM_3032580 (Figure 2C).
case_images: The effect of variant c.226+1G>A on the splicing of HES7 mRNA. (A) Construction of minigene plasmids. (B) Agarose electrophoresis results of RT-PCR products. M, DNA marker DL5000; WT, 293 T cells transfected with wild-type plasmids; MT, 293 T cells transfected with mutant plasmids. (C) Sanger sequencing of gel-purified products. (D) Effect of variant c.226+1G>A on the splicing of HES7 mRNA.
case_images: fped-11-1201999-g003.jpg
case_images: PMC10485611_01_fped-11-1201999-g003.jpg
case_images: F3
case_images: T cells (Figure 3A). The wild-type DNA fragments were amplified using genomic DNA from healthy controls according to the manufacturer's instructions. Electrophoresis results revealed that the full-length amplification product was 295 bp in 293 T cells transfected with the wild-type minigene plasmids, while the amplification products were 600 bp in 293 T cells transfected with the mutant plasmids (Figure 3B). However, the RT-PCR products of the mutant minigene plasmids also contained a complete intron3 sequence (Figure 3C), which proved that the variant HES7 c.226+1G>A resulted in the retention of intron3 in the mutant mRNA (PVS1_strong, Figure 3D).