Exon skipping induced by CRISPR-directed gene editing regulates the response to chemotherapy in non-small cell lung carcinoma cells.
Exon skipping induced by CRISPR-directed gene editing regulates the response to chemotherapy in non-small cell lung carcinoma cells.
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DOI：
10.1038/s41434-022-00324-7
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发表时间：
2022-06
期刊：
影响因子：
5.1
通讯作者：
中科院分区：
文献类型：
--
作者：
我们一直在开发CRISPR指导的基因编辑作为通过核因子红细胞2相关因子2（NRF 2）的遗传破坏治疗非小细胞肺癌（NSCLC）的增强疗法。NRF2促进肿瘤细胞存活以响应治疗干预，因此其失能应恢复或增强有效的药物作用。在这里，我们报告了NRF2破坏如何以CRISPR介导的外显子跳跃的形式导致附带损害。转录本和截短蛋白质的异质性群体对化疗产生可变的反应，这取决于哪个功能域缺失。我们鉴定和表征预测和未预测的转录本群体，并发现几种类型的转录本是通过外显子跳跃产生的，其中一个或两个NRF2外显子缺失。在一种特定情况下，单个核苷酸的存在或不存在决定了外显子是否通过重组外显子剪接增强子（ESE）而被跳过。我们分离并表征了NSCLC肿瘤细胞群中由CRISPR活性诱导的克隆的多样性，这是这项令人兴奋的技术的一个关键且经常被忽视的遗传副产品。最后，gRNA必须小心设计，以避免改变基因表达模式，这可能导致对实体瘤治疗的可变反应。
We have been developing CRISPR-directed gene editing as an augmentative therapy for the treatment of non-small cell lung carcinoma (NSCLC) by genetic disruption of Nuclear Factor Erythroid 2-Related Factor 2 (NRF2). NRF2 promotes tumor cell survival in response to therapeutic intervention and thus its disablement should restore or enhance effective drug action. Here, we report how NRF2 disruption leads to collateral damage in the form of CRISPR-mediated exon skipping. Heterogeneous populations of transcripts and truncated proteins produce a variable response to chemotherapy, dependent on which functional domain is missing. We identify and characterize predicted and unpredicted transcript populations and discover that several types of transcripts arise through exon skipping; wherein one or two NRF2 exons are missing. In one specific case, the presence or absence of a single nucleotide determines whether an exon is skipped or not by reorganizing Exonic Splicing Enhancers (ESEs). We isolate and characterize the diversity of clones induced by CRISPR activity in a NSCLC tumor cell population, a critical and often overlooked genetic byproduct of this exciting technology. Finally, gRNAs must be designed with care to avoid altering gene expression patterns that can account for variable responses to solid tumor therapy.
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DOI：
10.1016/j.omto.2018.10.002
发表时间：
2018-12-21
期刊：
Molecular therapy oncolytics
影响因子：
--
作者：
Bialk P;Wang Y;Banas K;Kmiec EB
通讯作者：
Kmiec EB
影响因子：
3.7
作者：
Lalonde S;Stone OA;Lessard S;Lavertu A;Desjardins J;Beaudoin M;Rivas M;Stainier DYR;Lettre G
通讯作者：
Lettre G
影响因子：
3
作者：
Chen, Jianli;Solomides, Charalambos;Simpkins, Henry
通讯作者：
Simpkins, Henry
影响因子：
4.6
作者：
Ono R;Ishii M;Fujihara Y;Kitazawa M;Usami T;Kaneko-Ishino T;Kanno J;Ikawa M;Ishino F
通讯作者：
Ishino F
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F