基因编辑升级!利用CRISPR-Cas9技术制造大范围染色体缺失
臂级染色体缺失是癌症的普遍且定义特征。 高度的肿瘤类型和亚型特异性复发提示选择性的致癌优势。 但是,由于它们的大尺寸,很难查明赋予这种优势的致癌驱动因子。 研究臂级缺失的致癌驱动能力的合适的功能基因组学方法是有限的。 在这里,我们介绍了一种有效的技术,可通过CRISPR-Cas9设计臂级缺失并创建同基因细胞系模型。 我们同时在染色体臂的两端诱导双链断裂(DSBs),并选择丢失了间歇区域的细胞。 使用这种技术,我们诱导神经母细胞瘤细胞系中的染色体11q(65 MB)和染色体6q(53 MB)的臂级缺失。 这种等基因模型使得能够进一步研究臂水平缺失在肿瘤发展和生长中的作用及其可能的治疗潜力。
Arm-level chromosomal deletions are a prevalent and defining feature of cancer. A high degree of tumor-type and subtype specific recurrencies suggest a selective oncogenic advantage. However, due to their large size it has been difficult to pinpoint the oncogenic drivers that confer this advantage. Suitable functional genomics approaches to study the oncogenic driving capacity of arm-level deletions are limited. Here we present an effective technique to engineer arm-level deletions by CRISPR-Cas9 and create isogenic cell line models. We simultaneously induce double-strand breaks (DSBs) at two ends of a chromosomal arm and select the cells that have lost the intermittent region. Using this technique, we induce arm-level deletions on chromosome 11q (65 MB) and chromosome 6q (53 MB) in neuroblastoma cell lines. Such isogenic models enable further research on the role of arm-level deletions in tumor development and growth and their possible therapeutic potential.
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