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September 2023 - Joana Ferreira da Silva and Goncalo Oliveira

Dr. Joana Ferreira da Silva and Dr. Gonçalo Oliveira

MedUni Wien RESEARCHER OF THE MONTH September 2023

Prime editing (PE) is a powerful genome engineering approach that enables the introduction of base substitutions, insertions and deletions into any given genomic locus. However, the efficiency of PE varies widely and depends not only on the genomic region targeted, but also on the genetic background of the edited cell. Here, to determine which cellular factors affect PE efficiency, we carry out a focused genetic screen targeting 32 DNA repair factors, spanning all reported repair pathways. We show that, depending on cell line and type of edit, ablation of mismatch repair (MMR) affords a 2-17 fold increase in PE efficiency, across several human cell lines, types of edits and genomic loci. The accumulation of the key MMR factors MLH1 and MSH2 at PE sites argues for direct involvement of MMR in PE control. Our results shed new light on the mechanism of PE and suggest how its efficiency might be optimised.

Selected Literature

  1. Ferreira da Silva, J.*, Oliveira, G.P*, Arasa-Verge, E.A., Kagiou, C., Moretton, A., Timelthaler, G., Jiricny, J. & Loizou, J.I. Prime editing efficiency and fidelity are enhanced in the absence of mismatch repair. Nature Communications 13, 760 (2022) *geteilte ErstautorInnen

  2. Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J.A. & Charpentier, E. A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science 337, 816-821 (2012)

  3. Ferreira da Silva, J., Salic, S., Wiedner, M., Datlinger, P., Essletzbichler, P., Hanzl, A., Superti-Furga, G., Bock, C., Loizou, J.I. Genome-scale CRISPR screens are efficient in non-homologous end-joining deficient cells. Scientific Reports 9, 15751 (2019)

  4. Anzalone, A.V., Randolph, P.B., Davis, J.R., Sousa, A.A., Koblan, L.W., Levy, J.M., Chen, P.J., Wilson, C., Newby, G.A., Raguram, A., Liu, D. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature 576, 149–157 (2019)

Dr. Joana Ferreira da Silva und Dr. Gonçalo Oliveira

Center for Genomic Medicine
Massachusetts General Hospital
Simches Research Building
185 Cambridge Street, Boston, MA 02114, US

Institute of Molecular Genetics of the Czech Academy of Sciences
Vídeňská 1083
142 20 Prague 4
Czech Republic