Efficient prime editing in two-cell mouse embryos using PEmbryo

Rebecca P Kim-Yip; Ryan McNulty; Bradley Joyce; Antonio Mollica; Peter J Chen; Purnima Ravisankar; Benjamin K Law; David R Liu; Jared E Toettcher; Evgueni A Ivakine; Eszter Posfai; Britt Adamson

doi:10.1038/s41587-023-02106-x

Efficient prime editing in two-cell mouse embryos using PEmbryo

Nat Biotechnol. 2024 Dec;42(12):1822-1830. doi: 10.1038/s41587-023-02106-x. Epub 2024 Feb 6.

Authors

Rebecca P Kim-Yip^#¹, Ryan McNulty^#², Bradley Joyce¹, Antonio Mollica^{3

4}, Peter J Chen^{5

6

7

8}, Purnima Ravisankar^{2

9}, Benjamin K Law^{1

2}, David R Liu^{5

6

7}, Jared E Toettcher¹, Evgueni A Ivakine^{3

10

11}, Eszter Posfai¹², Britt Adamson^{13

14}

Affiliations

¹ Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
² Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.
³ Genetics and Genome Biology Program, The Hospital for Sick Children, Toronto, ON, Canada.
⁴ Department of Biochemistry, University of Toronto, Toronto, ON, Canada.
⁵ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁶ Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
⁷ Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.
⁸ Prime Medicine, Inc., Cambridge, MA, USA.
⁹ Immunology and Microbial Pathogenesis Program, Weill Cornell Graduate School of Medical Sciences, New York, NY, USA.
¹⁰ Department of Physiology, University of Toronto, Toronto, ON, Canada.
¹¹ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
¹² Department of Molecular Biology, Princeton University, Princeton, NJ, USA. [email protected].
¹³ Department of Molecular Biology, Princeton University, Princeton, NJ, USA. [email protected].
¹⁴ Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA. [email protected].

^# Contributed equally.

Abstract

Using transient inhibition of DNA mismatch repair during a permissive stage of development, we demonstrate highly efficient prime editing of mouse embryos with few unwanted, local byproducts (average 58% precise edit frequency, 0.5% on-target error frequency across 13 substitution edits at 8 sites), enabling same-generation phenotyping of founders. Whole-genome sequencing reveals that mismatch repair inhibition increases off-target indels at low-complexity regions in the genome without any obvious phenotype in mice.

MeSH terms

Animals
CRISPR-Cas Systems / genetics
DNA Mismatch Repair / genetics
Embryo, Mammalian* / cytology
Female
Gene Editing* / methods
Mice

Abstract

MeSH terms

Grants and funding