ß-Globin Lentiviral Vectors Have Reduced Titers due to Incomplete Vector RNA Genomes and Lowered Virion Production.
Stem Cell Reports
; 16(1): 198-211, 2021 01 12.
Article
en En
| MEDLINE
| ID: mdl-33186538
ABSTRACT
Lentiviral vectors (LVs) commonly used for the treatment of hemoglobinopathies often have low titers and sub-optimal gene transfer efficiency for human hematopoietic stem and progenitor cells (HSPCs), hindering clinical translation and commercialization for ex vivo gene therapy. We observed that a high percentage of ß-globin LV viral genomic RNAs were incomplete toward the 3' end in packaging cells and in released vector particles. The incomplete vector genomes impeded reverse transcription in target cells, limiting stable gene transfer to HSPCs. By combining three modifications to vector design and production (shortening the vector length to 5.3 kb; expressing HIV-1 Tat protein during packaging; and packaging in PKR-/- cells) there was a 30-fold increase in vector titer and a 3-fold increase in vector infectivity in HSPCs. These approaches may improve the manufacturing of ß-globin and other complex LVs for enhanced gene delivery and may facilitate clinical applications.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Virión
/
ARN
/
Lentivirus
/
Globinas beta
/
Vectores Genéticos
Límite:
Humans
Idioma:
En
Revista:
Stem Cell Reports
Año:
2021
Tipo del documento:
Article
País de afiliación:
Estados Unidos