- Titel
- Dual Promoters Improve the Rescue of Recombinant Measles Virus in Human Cells
- Zitierfähige Url:
- https://nbn-resolving.org/urn:nbn:de:bsz:15-qucosa2-854124
- Quellenangabe
- Viruses
Erscheinungsjahr: 2021
Jahrgang: 13
Heft: 9
E-ISSN: 1999-4915
Artikelnummer: 1723 - Erstveröffentlichung
- 2021
- Abstract (EN)
- Reverse genetics is a technology that allows the production of a virus from its complementary DNA (cDNA). It is a powerful tool for analyzing viral genes, the development of novel vaccines, and gene delivery vectors. The standard reverse genetics protocols are laborious, time-consuming, and inefficient for negative-strand RNA viruses. A new reverse genetics platform was established, which increases the recovery efficiency of the measles virus (MV) in human 293-3-46 cells. The novel features compared with the standard system involving 293-3-46 cells comprise (a) dual promoters containing the RNA polymerase II promoter (CMV) and the bacteriophage T7 promoter placed in uni-direction on the same plasmid to enhance RNA transcription; (b) three G nucleotides added just after the T7 promoter to increase the T7 RNA polymerase activity; and (c) two ribozymes, the hairpin hammerhead ribozyme (HHRz), and the hepatitis delta virus ribozyme (HDVrz), were used to cleavage the exact termini of the antigenome RNA. Full-length antigenome cDNA of MV of the wild type IC323 strain or the vaccine AIK-C strain was inserted into the plasmid backbone. Both virus strains were easily rescued from their respective cloned cDNA. The rescue efficiency increased up to 80% compared with the use of the standard T7 rescue system. We assume that this system might be helpful in the rescue of other human mononegavirales.
- Andere Ausgabe
- Link zur Erstveröffentlichung
Link: https://doi.org/10.3390/v13091723 - Freie Schlagwörter (EN)
- measles virus; reverse genetics; negative-strand RNA virus; recombinant virus; multivalent vaccines; oncolytic vectors; mononegavirales
- Klassifikation (DDC)
- 610
- Verlag
- MDPI, Basel
- Version / Begutachtungsstatus
- publizierte Version / Verlagsversion
- URN Qucosa
- urn:nbn:de:bsz:15-qucosa2-854124
- Veröffentlichungsdatum Qucosa
- 09.05.2023
- Dokumenttyp
- Artikel
- Sprache des Dokumentes
- Englisch
- Lizenz / Rechtehinweis
CC BY 4.0