Fig. 3
From: Novel MNX1 mutations and genotype–phenotype analysis of patients with Currarino syndrome
Minigene study on a recurrent noncanonical splice site variant in MNX1. a Structure of the splicing vector pEGFP-C1 and minigene MNX1-wt/MNX1-mut (c.691 + 3G > T): the pEGFP-C1 vector contains a CMV promoter, and the symbol “*” represents the location of the mutation. b Sequencing results of the target fragment with wild-type (wt) at the top and mutant (mut) at the bottom. c Reverse-transcription polymerase chain reaction (RT-PCR) products were separated by electrophoresis in HEK-293 T (left) and HeLa (right) cells. d Minigene product sequencing results: a, the wild-type minigene (MNX1-wt) formed normal mRNA composed of exons 1 and 2; b, the mutant intron c.691 + 3G > T minigene caused a splicing abnormality in HeLa cells, resulting in the skipping of the 147 bp base in exon 1