Frameshift indels introduced by genome editing can lead to in-frame exon skipping.
Frameshift indels introduced by genome editing can lead to in-frame exon skipping.
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DOI：
10.1371/journal.pone.0178700
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Lettre G
中科院分区：
文献类型：
--
作者：
Lalonde S;Stone OA;Lessard S;Lavertu A;Desjardins J;Beaudoin M;Rivas M;Stainier DYR;Lettre G
通过基因组编辑引入移码插入缺失已经成为研究细胞系和模式生物中未表征基因功能的强大技术。这种突变会导致mRNA降解，这是由于无义介导的mRNA降解或产生严重截短的蛋白质。在这里，我们表明，通过基因组编辑工程化的移码插入缺失也可以导致“三个核苷酸的倍数”外显子的跳跃。这样的剪接事件导致可以编码完全或部分功能蛋白质的框内mRNA。我们还表征了位于LGALS 8的“三个核苷酸的倍数”外显子中的分离无义变体（rs2273865），其增加了人类成红细胞样品中的外显子跳跃。我们的研究结果突出了外显子剪接调控元件的潜在频繁贡献，对于解释基因组编辑实验中的阴性结果非常重要。此外，它们可能有助于更好地注释人类基因组中的功能丧失突变。
The introduction of frameshift indels by genome editing has emerged as a powerful technique to study the functions of uncharacterized genes in cell lines and model organisms. Such mutations should lead to mRNA degradation owing to nonsense-mediated mRNA decay or the production of severely truncated proteins. Here, we show that frameshift indels engineered by genome editing can also lead to skipping of “multiple of three nucleotides” exons. Such splicing events result in in-frame mRNA that may encode fully or partially functional proteins. We also characterize a segregating nonsense variant (rs2273865) located in a “multiple of three nucleotides” exon of LGALS8 that increases exon skipping in human erythroblast samples. Our results highlight the potentially frequent contribution of exonic splicing regulatory elements and are important for the interpretation of negative results in genome editing experiments. Moreover, they may contribute to a better annotation of loss-of-function mutations in the human genome.