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. 2021 Jan 11;49(3):1769–1783. doi: 10.1093/nar/gkaa1167

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Neochromosomes as landing pads. (A) 35 kb of non-coding DNA was simultaneously in vivo assembled and inserted by CRISPR/Cas9 at the mTurquoise2 locus of the 100 kb neochromosome in IMF6 and of the 50 kb neochromosome in IMF2, resulting in 135 kb (IMF11) and 85 kb (IMF12) neochromosomes, respectively. (B) 35 kb of glycolytic genes were simultaneously in vivo assembled and inserted using CRISPR/Cas9 at the mTurquoise2 locus of the 100 kb neochromosome in IMF6 and of the 50 kb neochromosome in IMF2 resulting in 135 kb (IMF13) and 85 kb (IMF14) neochromosomes, respectively. As control, the same glycolytic cassettes were integrated at the native CAN1 locus of chromosome V resulting in strain IMX1959.