Engineered virus-like particles for transient delivery of prime editor ribonucleoprotein complexes in vivo

Meirui An; Aditya Raguram; Samuel W Du; Samagya Banskota; Jessie R Davis; Gregory A Newby; Paul Z Chen; Krzysztof Palczewski; David R Liu

doi:10.1038/s41587-023-02078-y

Engineered virus-like particles for transient delivery of prime editor ribonucleoprotein complexes in vivo

Nat Biotechnol. 2024 Jan 8. doi: 10.1038/s41587-023-02078-y. Online ahead of print.

Authors

Meirui An^{1

2

3}, Aditya Raguram^{1

2

3}, Samuel W Du^{4

5}, Samagya Banskota^{1

2

3}, Jessie R Davis^{1

2

3}, Gregory A Newby^{1

2

3}, Paul Z Chen^{1

2

3}, Krzysztof Palczewski^{4

5

6

7}, David R Liu^{8

9

10}

Affiliations

¹ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
² Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
³ Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.
⁴ Gavin Herbert Eye Institute, Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, CA, USA.
⁵ Department of Physiology and Biophysics, University of California, Irvine, CA, USA.
⁶ Department of Chemistry, University of California, Irvine, CA, USA.
⁷ Department of Molecular Biology and Biochemistry, University of California, Irvine, CA, USA.
⁸ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA. drliu@fas.harvard.edu.
⁹ Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA. drliu@fas.harvard.edu.
¹⁰ Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA. drliu@fas.harvard.edu.

PMID: 38191664
DOI: 10.1038/s41587-023-02078-y

Abstract

Prime editing enables precise installation of genomic substitutions, insertions and deletions in living systems. Efficient in vitro and in vivo delivery of prime editing components, however, remains a challenge. Here we report prime editor engineered virus-like particles (PE-eVLPs) that deliver prime editor proteins, prime editing guide RNAs and nicking single guide RNAs as transient ribonucleoprotein complexes. We systematically engineered v3 and v3b PE-eVLPs with 65- to 170-fold higher editing efficiency in human cells compared to a PE-eVLP construct based on our previously reported base editor eVLP architecture. In two mouse models of genetic blindness, single injections of v3 PE-eVLPs resulted in therapeutically relevant levels of prime editing in the retina, protein expression restoration and partial visual function rescue. Optimized PE-eVLPs support transient in vivo delivery of prime editor ribonucleoproteins, enhancing the potential safety of prime editing by reducing off-target editing and obviating the possibility of oncogenic transgene integration.