Julien Capin
An engineered baculoviral protein and DNA co-delivery system for CRISPR-based mammalian genome editing
Capin, Julien; Harrison, Alexandra; Raele, Renata A.; Yadav, Sathish K.N.; Baiwir, Dominique; Mazzucchelli, Gabriel; Quinton, Loic; Satchwell, Timothy J.; Toye, Ashley M.; Schaffitzel, Christiane; Berger, Imre; Aulicino, Francesco
Authors
Alexandra Harrison
Renata A. Raele
Sathish K.N. Yadav
Dominique Baiwir
Gabriel Mazzucchelli
Loic Quinton
Timothy J. Satchwell
Ashley M. Toye
Christiane Schaffitzel
Imre Berger
Francesco Aulicino
Abstract
CRISPR-based DNA editing technologies enable rapid and accessible genome engineering of eukaryotic cells. However, the delivery of genetically encoded CRISPR components remains challenging and sustained Cas9 expression correlates with higher off-target activities, which can be reduced via Cas9-protein delivery. Here we demonstrate that baculovirus, alongside its DNA cargo, can be used to package and deliver proteins to human cells. Using protein-loaded baculovirus (pBV), we demonstrate delivery of Cas9 or base editors proteins, leading to efficient genome and base editing in human cells. By implementing a reversible, chemically inducible heterodimerization system, we show that protein cargoes can selectively and more efficiently be loaded into pBVs (spBVs). Using spBVs we achieved high levels of multiplexed genome editing in a panel of human cell lines. Importantly, spBVs maintain high editing efficiencies in absence of detectable off-targets events. Finally, by exploiting Cas9 protein and template DNA co-delivery, we demonstrate up to 5% site-specific targeted integration of a 1.8 kb heterologous DNA payload using a single spBV in a panel of human cell lines. In summary, we demonstrate that spBVs represent a versatile, efficient and potentially safer alternative for CRISPR applications requiring co-delivery of DNA and protein cargoes.
Journal Article Type | Article |
---|---|
Acceptance Date | Feb 15, 2024 |
Online Publication Date | Feb 27, 2024 |
Publication Date | Apr 12, 2024 |
Deposit Date | Jul 11, 2024 |
Publicly Available Date | Jul 12, 2024 |
Journal | Nucleic Acids Research |
Print ISSN | 0305-1048 |
Publisher | Oxford University Press (OUP) |
Peer Reviewed | Peer Reviewed |
Volume | 52 |
Issue | 6 |
Pages | 3450-3468 |
DOI | https://doi.org/10.1093/nar/gkae142 |
Public URL | https://uwe-repository.worktribe.com/output/12120130 |
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An engineered baculoviral protein and DNA co-delivery system for CRISPR-based mammalian genome editing
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Licence
http://creativecommons.org/licenses/by/4.0/
Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/
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