Genome editing is induced in a binary manner in single human cells

Gou Takahashi; Daiki Kondo; Minato Maeda; Yuji Morishita; Yuichiro Miyaoka

doi:10.1016/j.isci.2022.105619

Genome editing is induced in a binary manner in single human cells

iScience. 2022 Nov 17;25(12):105619. doi: 10.1016/j.isci.2022.105619. eCollection 2022 Dec 22.

Authors

Gou Takahashi¹, Daiki Kondo^{1

2}, Minato Maeda^{1

2}, Yuji Morishita³, Yuichiro Miyaoka^{1

2

4}

Affiliations

¹ Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
² Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
³ On-chip Biotechnologies Co., Ltd., Tokyo, Japan.
⁴ Graduate School of Humanities and Sciences, Ochanomizu University, Tokyo, Japan.

Abstract

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.

Keywords: Bioengineering; Biological sciences; Biotechnology; Genetic engineering.