Cas9 protein delivery non-integrating lentiviral vectors for gene correction in sickle cell disease.
CRISPR-Cas9
gene correction
genome editing
lentiviral vector
non-integrating vector
protein delivery
sickle cell disease
Journal
Molecular therapy. Methods & clinical development
ISSN: 2329-0501
Titre abrégé: Mol Ther Methods Clin Dev
Pays: United States
ID NLM: 101624857
Informations de publication
Date de publication:
11 Jun 2021
11 Jun 2021
Historique:
received:
22
02
2021
accepted:
26
02
2021
entrez:
5
4
2021
pubmed:
6
4
2021
medline:
6
4
2021
Statut:
epublish
Résumé
Gene editing with the CRISPR-Cas9 system could revolutionize hematopoietic stem cell (HSC)-targeted gene therapy for hereditary diseases, including sickle cell disease (SCD). Conventional delivery of editing tools by electroporation limits HSC fitness due to its toxicity; therefore, efficient and non-toxic delivery remains crucial. Integrating lentiviral vectors are established for therapeutic gene delivery to engraftable HSCs in gene therapy trials; however, their sustained expression and size limitation preclude their use for CRISPR-Cas9 delivery. Here, we developed a Cas9 protein delivery non-integrating lentiviral system encoding guide RNA and donor DNA, allowing for transient endonuclease function and inclusion of all editing tools in a single vector (all-in-one). We demonstrated efficient one-time correction of the SCD mutation in the endogenous βs-globin gene up to 42% at the protein level (p < 0.01) with the Cas9 protein delivery non-integrating lentiviral all-in-one system without electroporation. Our findings improve prospects for efficient and safe genome editing.
Identifiants
pubmed: 33816645
doi: 10.1016/j.omtm.2021.02.022
pii: S2329-0501(21)00037-1
pmc: PMC8005818
doi:
Types de publication
Journal Article
Langues
eng
Pagination
121-132Déclaration de conflit d'intérêts
The authors declare no competing interests.
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