Genome Editing Is Induced in a Binary Manner in Single Human Cells
Genome Editing Is Induced in a Binary Manner in Single Human Cells
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基因组编辑在单个人类细胞中以二元方式诱导
DOI：
10.1101/2022.03.04.482947
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Miyaoka Yuichiro
中科院分区：
文献类型：
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作者：
Takahashi Gou;Kondo Daiki;Maeda Minato;Morishita Yuji;Miyaoka Yuichiro
即使打算进行精确的核苷酸操作，基因组编辑的结果也可能是多种多样的，通常包括随机插入和缺失。这些不同结果在单细胞中的组合和频率不仅在转基因细胞系的生成中至关重要，而且在评估基因组编辑疗法的临床效果中也至关重要。然而，目前的方法仅分析细胞群体，而不是单个细胞。在这里，我们利用单粒子分离系统（SPiS）有效分离单细胞，系统地分析单个人类培养细胞中的基因组编辑结果。结果，我们发现基因组编辑诱导具有二元性质，即细胞的靶等位基因倾向于全部被编辑或根本不被编辑。这项研究增强了我们对基因组编辑诱导模式的理解，并提供了一种新的策略来分析单细胞中的基因组编辑结果。
Even when precise nucleotide manipulations are intended, the outcomes of genome editing can be diverse, often including random insertions and deletions. The combinations and frequencies of these different outcomes in single cells are critical not only in the generation of genetically modified cell lines but also in the evaluation of the clinical effects of genome editing therapies. However, current methods only analyze cell populations, not single cells. Here, we utilized the Single Particle isolation System (SPiS) for the efficient isolation of single cells to systematically analyze genome editing results in individual human cultured cells. As a result, we discovered that genome editing induction has a binary nature, that is, the target alleles of cells tend to be all edited or not edited at all. This study enhances our understanding of the induction pattern of genome editing and provides a new strategy to analyze genome editing outcomes in single cells.