Genome editing in human iPS cells to study and cure disease

 

Yuichiro Miyaoka, PhD.1,2,3

1 Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science

2 Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University

3 Graduate School of Humanities and Sciences, Ochanomizu University

 

Abstract

Genome editing technology allows us to rewrite the genetic information in virtually any species and any cell type including human cells. Genome editing in human iPS cells is particularly useful in studying and curing disease. Therefore, we established a digital PCR-based platform to detect and isolate genome-edited human iPS cells without using any selection marker genes [1,2]. By using this platform, we have established isogenic iPS cell-based models and elucidated the pathogenic mechanisms of cardiomyopathy and liver disease [3,4]. We have also proven that genome-edited iPS cells can be utilized to deliver therapeutic molecules in vivo [5].

We have also been working on improving the efficiency of isolation of genome-edited cells. For this purpose, it is very important to know genome editing outcomes in single cells. Therefore, we have developed a system for large-scale single-cell cloning of human cultured cells and iPS cells to analyze the genome editing results individually [6,7].

Today, I would like to present the recent progress in our lab to study and cure genetic disorders by genome editing in human iPS cells.

 

References

1 Miyaoka Y, Chan AH, Judge LM, Yoo J, Huang M, Nguyen TD, Lizarraga PP, So P-L & Conklin BR (2014) Isolation of single-base genome-edited human iPS cells without antibiotic selection. Nat. Methods 11, 291–293.

2 *#Miyaoka Y, *Berman JR, Cooper SB, Mayerl SJ, Chan AH, Zhang B, Karlin-Neumann GA & #Conklin BR (2016) Systematic quantification of HDR and NHEJ reveals effects of locus, nuclease, and cell type on genome-editing. Sci. Rep. 6, 23549.

3 *Song D, *Takahashi G, Zheng Y-W, Matsuo-Takasaki M, Li J, Takami M, An Y, Hemmi Y, Miharada N, Fujioka T, Noguchi M, Nakajima T, Saito MK, Nakamura Y, Oda T, #Miyaoka Y & #Hayashi Y (2022) Retinoids rescue ceruloplasmin secretion and alleviate oxidative stress in Wilson’s disease-specific hepatocytes. Hum. Mol. Genet. 31, 3652–3671.

4 *Fenix AM, *Miyaoka Y, Bertero A, Blue SM, Spindler MJ, Tan KKB, Perez-Bermejo JA, Chan AH, Mayerl SJ, Nguyen TD, Russell CR, Lizarraga PP, Truong A, So P-L, Kulkarni A, Chetal K, Sathe S, Sniadecki NJ, Yeo GW, #Murry CE, #Conklin BR & #Salomonis N (2021) Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein granules within processing bodies. Nat. Commun. 12, 6324.

5 Nakajima I, Tsukimura T, Ono T, Shiga T, Shitara H, Togawa T, Sakuraba H & Miyaoka Y (2023) In Vivo Delivery of Therapeutic Molecules by Transplantation of Genome-Edited Induced Pluripotent Stem Cells. Cell Transplant. 32, 9636897231173734.

6 Takahashi G & Miyaoka Y (2023) Large-scale single-cell cloning of genome-edited cultured human cells by On-chip SPiS. STAR Protocols 4, 102364.

7 Takahashi G, Kondo D, Maeda M, Morishita Y & Miyaoka Y (2022) Genome editing is induced in a binary manner in single human cells. iScience 25, 105619.