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

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.