Enabling large-scale genome editing at repetitive elements by reducing DNA nicking
Research output: Contribution to journal › Journal article › Research › peer-review
To extend the frontier of genome editing and enable editing of repetitive elements of mammalian genomes, we made use of a set of dead-Cas9 base editor (dBE) variants that allow editing at tens of thousands of loci per cell by overcoming the cell death associated with DNA double-strand breaks and single-strand breaks. We used a set of gRNAs targeting repetitive elements-ranging in target copy number from about 32 to 161 000 per cell. dBEs enabled survival after large-scale base editing, allowing targeted mutations at up to similar to 13 200 and similar to 12 200 loci in 293T and human induced pluripotent stem cells (hiP-SCs), respectively, three orders of magnitude greater than previously recorded. These dBEs can overcome current on-target mutation and toxicity barriers that prevent cell survival after large-scale genome engineering.
Original language | English |
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Journal | Nucleic Acids Research |
Volume | 48 |
Issue number | 9 |
Pages (from-to) | 5183-5195 |
Number of pages | 13 |
ISSN | 0305-1048 |
DOIs | |
Publication status | Published - 21 May 2020 |
Externally published | Yes |
- L1 RETROTRANSPOSITION, STEM-CELLS, BASE, ORGANISMS, MUTATION, REPAIR, MOBILE
Research areas
ID: 257032520