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bioRxiv · 10.1101/342311

\"Ancestralization\" of human pluripotent stem cells by multiplexed precise genome editing

Riesenberg, S.·Maricic, T.·Pääbo, S.
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Abstract

We show that inactivation of the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) results in a drastic increase in efficiency of precise genome editing with CRISPR enzymes in human stem cells, allowing up to 79% of chromosomes to carry an intended nucleotide substitution when a single genomic site is targeted. When three different genes are simultaneously targeted, 12% of the isolated cells carry the targeted amino acid-changing substitutions in homozygous forms. These substitutions represent the first step towards resurrecting the proteome ancestral to Neandertals and modern humans. DNA-PKcs inactivation will greatly facilitate multiplexed precise genome editing in animal cells.

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Riesenberg, S., Maricic, T., Pääbo, S.. 2018-06-08. \"Ancestralization\" of human pluripotent stem cells by multiplexed precise genome editing. https://doi.org/10.1101/342311

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