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Genome Editing Is Induced in a Binary Manner in Single Human Cells

Gou Takahashi, Daiki Kondo, Minato Maeda, Yuji Morishita, View ORCID ProfileYuichiro Miyaoka
doi: https://doi.org/10.1101/2022.03.04.482947
Gou Takahashi
1Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
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Daiki Kondo
1Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
2Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
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Minato Maeda
1Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
3Department of Biological Sciences, Faculty of Science and Engineering, Chuo University, Tokyo, Japan
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Yuji Morishita
4On-chip Biotechnologies Co., Ltd., Tokyo, Japan
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Yuichiro Miyaoka
1Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
2Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
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  • ORCID record for Yuichiro Miyaoka
  • For correspondence: miyaoka-yi@igakuken.or.jp
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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 mechanism of genome editing and provides a new strategy to analyze genome editing outcomes in single cells.

Competing Interest Statement

Y.Morishita is an employee of On-chip Biotechnologies Co., Ltd.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted March 05, 2022.
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Genome Editing Is Induced in a Binary Manner in Single Human Cells
Gou Takahashi, Daiki Kondo, Minato Maeda, Yuji Morishita, Yuichiro Miyaoka
bioRxiv 2022.03.04.482947; doi: https://doi.org/10.1101/2022.03.04.482947
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Genome Editing Is Induced in a Binary Manner in Single Human Cells
Gou Takahashi, Daiki Kondo, Minato Maeda, Yuji Morishita, Yuichiro Miyaoka
bioRxiv 2022.03.04.482947; doi: https://doi.org/10.1101/2022.03.04.482947

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