Genes adapt to outsmart gene-targeting strategies in mutant mouse strains by skipping exons to reinitiate transcription and translation
Genes adapt to outsmart gene-targeting strategies in mutant mouse strains by skipping exons to reinitiate transcription and translation
复制标题
DOI：
10.1186/s13059-020-02086-0
复制
发表时间：
2020-07-09
期刊：
影响因子：
12.3
通讯作者：
Wiles, Michael, V
中科院分区：
文献类型：
--
作者：
Hosur, Vishnu;Low, Benjamin E.;Wiles, Michael, V
小鼠胚胎干细胞或受精卵的基因破坏是鉴定体内基因功能的传统遗传学方法。然而，由于不同的基因破坏策略使用不同的机制来破坏基因，这些策略可以在所得到的小鼠模型中导致不同的表型。为了确定不同的基因破坏策略是否会影响突变小鼠的表型，我们对三种常用策略（明确无效、靶向敲除(KO)-first和CRISPR/Cas9）产生的drhbdf1小鼠突变株进行了表征。结果我们发现rhbdf1对不同的KO策略有不同的反应，例如，通过跳过外显子和重新启动翻译来产生潜在的功能获得等位基因，而不是预期的零等位基因或严重的次形等位基因。我们的分析还显示，使用ko优先策略产生的小鼠中至少有4%表现出冲突的表型。结论外显子跳变是基因组中普遍存在的现象。这些发现对基因组编辑在基础研究和临床实践中的应用具有重要意义。
Background Gene disruption in mouse embryonic stem cells or zygotes is a conventional genetics approach to identify gene function in vivo. However, because different gene disruption strategies use different mechanisms to disrupt genes, the strategies can result in diverse phenotypes in the resulting mouse model. To determine whether different gene disruption strategies affect the phenotype of resulting mutant mice, we characterizedRhbdf1mouse mutant strains generated by three commonly used strategies-definitive-null, targeted knockout (KO)-first, and CRISPR/Cas9. Results We find thatRhbdf1responds differently to distinct KO strategies, for example, by skipping exons and reinitiating translation to potentially yield gain-of-function alleles rather than the expected null or severe hypomorphic alleles. Our analysis also revealed that at least 4% of mice generated using the KO-first strategy show conflicting phenotypes. Conclusions Exon skipping is a widespread phenomenon occurring across the genome. These findings have significant implications for the application of genome editing in both basic research and clinical practice.