Fig. 2: Novel splice defects due to deep-intronic ABCA4 variants. | Genetics in Medicine

Fig. 2: Novel splice defects due to deep-intronic ABCA4 variants.

From: Resolving the dark matter of ABCA4 for 1054 Stargardt disease probands through integrated genomics and transcriptomics

Fig. 2: Novel splice defects due to deep-intronic ABCA4 variants.

Wild-type (WT) and mutant (MT) midigenes were transfected in HEK293T cells and the extracted RNA was subjected to reverse-transcription polymerase chain reaction (RT-PCR). Left panels show the ABCA4-specific RT-PCR products with Rhodopsin exon 5 (RHO e5) RT-PCR as a transfection efficiency control. In the middle panels, Sanger sequencing results of the RT-PCR products are given. At the right side, pseudoexons (PEs) and an exon elongation are depicted with splice site strength predictions for WT and MT sequences, with green rectangles representing the splice acceptor sites and blue rectangles representing the splice donor sites. Red highlighted nucleotides represent the variants. Except for c.1937+37C>G (e), which resulted in a 36-nt exon 13 elongation, all deep-intronic variants lead to PEs (a–d, f, g). The intron 7 variants in (d) result in partially overlapping PEs that share the same splice acceptor site at position c.859-685. HSF Human Splicing Finder, na not applicable, PE pseudoexon, SSFL splice site finder–like.

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