Construction and Characterization of a High-Capacity Replication-Competent Murine Cytomegalovirus Vector for Gene Delivery

We investigated the basic characteristics of a new murine cytomegalovirus (MCMV) vector platform. Using BAC technology, we engineered replication-competent recombinant MCMVs with deletions of up to 26% of the wild-type genome. To this end, we targeted five gene blocks (m01-m17, m106-m109, m129-m141,...

Full description

Saved in:
Bibliographic Details
Published in:Vaccines (Basel) 2024-07, Vol.12 (7), p.791
Main Authors: Riedl, André, Bojková, Denisa, Tan, Jiang, Jeney, Ábris, Larsen, Pia-Katharina, Jeney, Csaba, Full, Florian, Kalinke, Ulrich, Ruzsics, Zsolt
Format: Article
Language:English
Subjects:
Analysis
Antigens
Artificial chromosomes
Cell lines
Cloning
Cloning vectors
cross-species application
Cytomegalovirus
Cytomegaloviruses
Dosage and administration
E coli
Fibroblasts
Gene transfer
gene transfer vector
Genes
Genetic aspects
genetic stability
Genetic vectors
Genomes
Genotype & phenotype
Herpes viruses
high-capacity vector
host range
Identification and classification
Infections
Inserts
Lymphocytes
Mammalian cells
Mammals
Pathogens
Phenotypes
Plasmids
Replication
Vaccines
Vectors (Biology)
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We investigated the basic characteristics of a new murine cytomegalovirus (MCMV) vector platform. Using BAC technology, we engineered replication-competent recombinant MCMVs with deletions of up to 26% of the wild-type genome. To this end, we targeted five gene blocks (m01-m17, m106-m109, m129-m141, m144-m158, and m159-m170). BACs featuring deletions from 18% to 26% of the wild-type genome exhibited delayed virus reconstitution, while smaller deletions (up to 16%) demonstrated reconstitution kinetics similar to those of the wild type. Utilizing an innovative methodology, we introduced large genomic DNA segments, up to 35 kbp, along with reporter genes into a newly designed vector with a potential cloning capacity of 46 kbp (Q4). Surprisingly, the insertion of diverse foreign DNAs alleviated the delayed plaque formation phenotype of Q4, and these large inserts remained stable through serial in vitro passages. With reporter-gene-expressing recombinant MCMVs, we successfully transduced not only mouse cell lines but also non-rodent mammalian cells, including those of human, monkey, bovine, and bat origin. Remarkably, even non-mammalian cell lines derived from chickens exhibited successful transduction.
ISSN:2076-393X
2076-393X
DOI:10.3390/vaccines12070791