Generation of Monosomy 21q Human iPS Cells by CRISPR/Cas9-Mediated Interstitial Megabase Deletion.

Autor: Egawa M; Laboratory of Bioengineering, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan., Uno N; Laboratory of Bioengineering, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan., Komazaki R; Laboratory of Bioengineering, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan., Ohkame Y; Chromosome Engineering Research Center, Tottori University, Tottori, Japan., Yamazaki K; Chromosome Engineering Research Group, the Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Okazaki, Japan.; Department of Homeostatic Regulation, National Institute for Physiological Sciences (NIPS), National Institutes of Natural Sciences (NINS), Okazaki, Japan., Yoshimatsu C; Department of Chromosome Biomedical Engineering, Integrated Medical Sciences, Graduate School of Medical Sciences, Tottori University, Tottori, Japan., Ishizu Y; Laboratory of Bioengineering, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan., Okano Y; Laboratory of Bioengineering, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan., Miyamoto H; Department of Chromosome Biomedical Engineering, Integrated Medical Sciences, Graduate School of Medical Sciences, Tottori University, Tottori, Japan., Osaki M; Chromosome Engineering Research Center, Tottori University, Tottori, Japan.; Division of Experimental Pathology, Faculty of Medicine, Tottori University, Tottori, Japan., Suzuki T; Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan., Hosomichi K; Laboratory of Computational Genomics, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan., Aizawa Y; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.; Kanagawa Institute of Industrial Science and Technology, Ebina, Japan., Kazuki Y; Chromosome Engineering Research Center, Tottori University, Tottori, Japan.; Chromosome Engineering Research Group, the Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Okazaki, Japan.; Department of Chromosome Biomedical Engineering, Integrated Medical Sciences, Graduate School of Medical Sciences, Tottori University, Tottori, Japan., Tomizuka K; Laboratory of Bioengineering, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan.
Jazyk: angličtina
Zdroj: Genes to cells : devoted to molecular & cellular mechanisms [Genes Cells] 2024 Nov 24. Date of Electronic Publication: 2024 Nov 24.
DOI: 10.1111/gtc.13184
Abstrakt: Missing an entire chromosome or chromosome arm in normal diploid cells has a deleterious impact on cell viability, which may contribute to the development of specific birth defects. Nevertheless, the effects of chromosome loss in human cells have remained unexplored due to the lack of suitable model systems. Here, we developed an efficient, selection-free approach to generate partial monosomy in human induced pluripotent stem cells (iPSCs). The introduction of Cas9 proteins and a pair of gRNAs induces over megabase-sized interstitial chromosomal deletions. Using human chromosome 21 (HSA21) as a model, partial monosomy 21q (PM21q) iPSC lines with deletions ranging from 4.5 to 27.9 Mb were isolated. A 33.6 Mb deletion, encompassing all protein-coding genes on 21q, was also achieved, establishing the first 21q monosomy human iPSC line. Transcriptome and proteome analyses revealed that the abundances of mRNA and protein encoded by the majority of genes in the monosomic regions are half of the diploid expression level, indicating an absence of dosage compensation. The ability to generate customized partial monosomy cell lines on an isogenic, karyotypically normal background should facilitate the gain of novel insights into the impact of chromosome loss on cellular fitness.
(© 2024 The Author(s). Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.)
Databáze: MEDLINE
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