CRISPR/Cas9-mediated genome editing induces exon skipping by complete or stochastic altering splicing in the migratory locust.
CRISPR/Cas9-mediated genome editing induces exon skipping by complete or stochastic altering splicing in the migratory locust.
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CRISPR/Cas9 介导的基因组编辑通过完全或随机改变剪接在飞蝗中诱导外显子跳跃
DOI：
10.1186/s12896-018-0465-7
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发表时间：
2018-09-25
影响因子：
3.5
通讯作者：
Kang L
中科院分区：
文献类型：
--
作者：
Chen D;Tang JX;Li B;Hou L;Wang X;Kang L
背景CRISPR/Cas9系统已被广泛应用于通过在细胞系和生物体中诱导移码突变来建立基因敲除/敲除模型。最近的几项研究报道，这种突变可以导致细胞系中的框内外显子跳跃。然而，关于CRISPR介导的基因编辑在体内的转录后效应的研究很少。结果我们发现，移码插入还通过删除不同的区域来影响体内的Pre-mRNA剪接，从而诱导完全或随机的外显子跳跃。在蝗虫中，嗅觉受体基因Lmior35内含子3和外显子4交界处缺失55个碱基，导致外显子4完全跳跃，而外显子4缺失22个碱基，仅导致随机外显子4跳跃。单个sgRNA诱导内含子和外显子交界处的小片段插入或缺失破坏3‘剪接点，导致完全外显子跳跃，或者诱导外显子小插入或小缺失导致随机改变剪接，导致随机外显子跳跃。结论CRISPR介导的基因组编辑通过删除基因的不同区域，可能导致完全或随机外显子跳跃。尽管CRISPR介导的编辑导致的外显子跳跃是一个意想不到的结果，但这一发现可以作为一种技术来研究前mRNA剪接或治疗由剪接突变引起的几种人类疾病。
BackgroundThe CRISPR/Cas9 system has been widely used to generate gene knockout/knockin models by inducing frameshift mutants in cell lines and organisms. Several recent studies have reported that such mutants can lead to in-frame exon skipping in cell lines. However, there was little research about post-transcriptional effect of CRISPR-mediated gene editing in vivo.ResultsWe showed that frameshift indels also induced complete or stochastic exon skipping by deleting different regions to influence pre-mRNA splicing in vivo. In the migratory locust, the missing 55 bp at the boundary of intron 3 and exon 4 of an olfactory receptor gene,LmigOr35, resulted in complete exon 4 skipping, whereas the lacking 22 bp in exon 4 ofLmigOr35only resulted in stochastic exon 4 skipping. A single sgRNA induced small insertions or deletions at the boundary of intron and exon to disrupt the 3′ splicing site causing completely exon skipping, or alternatively induce small insertions or deletions in the exon to stochastic alter splicing causing the stochastic exon skipping.ConclusionsThese results indicated that complete or stochastic exon skipping could result from the CRISPR-mediated genome editing by deleting different regions of the gene. Although exon skipping caused by CRISPR-mediated editing was an unexpected outcome, this finding could be developed as a technology to investigate pre-mRNA splicing or to cure several human diseases caused by splicing mutations.
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影响因子：
4.4
作者：
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通讯作者：
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64.5
作者：
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通讯作者：
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