A short sequence within AluSx induces downstream exon skipping in an ACAT1 minigene model

Abstract

Alu elements, the most abundant retrotransposed elements in the human genome, are commonly located in introns and affect alternative splicing. We previously showed that an intronic antisense Alu element enhanced alternative splicing in a minigene model. The RNA splicing experiment was performed using a minigene consisting of exons 9–11 of ACAT1 with an antisense AluSx inserted into intron 10 in a pCAGGS vector. Here, we explored the sequence in the intronic antisense AluSx that affects downstream exon skipping. Our RNA splicing analysis with antisense AluSx deletion mutants confirmed a short sequence of 16 bp within the right arm of antisense AluSx that resulted in exon skipping. Our findings help clarify the mechanism of RNA splicing by Alu elements.

KEYWORDS:

• Alu elements
• alternative splicing
• exon skipping
• ACAT1
• antisense orientation

Acknowledgments

We gratefully appreciate the late Professor Toshiyuki Fukao, who established the minigene splicing model of this research and gave us this research theme. We thank Gabrielle White Wolf, PhD, from Edanz Group (https://en-author-services.edanzgroup.com/ac) for editing a draft of this manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data that support the findings of this study are available at the figshare repository (https://figshare.com/) at https://doi.org/10.6084/m9.figshare.16531881.v1. Nucleotide sequences of antisense AluSx in this study are available at https://doi.org/10.6084/m9.figshare.16531659.