tetano
Editor, Senior Moderator
Gene Ther. 2014 May 15. doi: 10.1038/gt.2014.37. [Epub ahead of print]
Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines.
Walters AA1, Kinnear E1, Shattock RJ1, McDonald JU1, Caproni LJ2, Porter N2, Tregoning JS1.
Author information
Abstract
The use of DNA to deliver vaccine antigens offers many advantages, including ease of manufacture and cost. However, most DNA vaccines are plasmids and must be grown in bacterial culture, necessitating elements that are either unnecessary for effective gene delivery (for example, bacterial origins of replication) or undesirable (for example, antibiotic resistance genes). Removing these elements may improve the safety profile of DNA for the delivery of vaccines. Here, we describe a novel, double-stranded, linear DNA construct produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, polyA tail and telomeric ends. We compared these constructs (called 'Doggybones' because of their shape) with conventional plasmid DNA. Using luciferase-expressing constructs, we demonstrated that expression levels were equivalent between Doggybones and plasmids both in vitro and in vivo. When mice were immunized with DNA constructs expressing the HIV envelope protein gp140, equivalent humoral and cellular responses were induced. Immunizations with either construct type expressing hemagluttinin were protective against H1N1 influenza challenge. This is the first example of an effective DNA vaccine, which can be produced on a large scale by enzymatic processes.Gene Therapy advance online publication, 15 May 2014; doi:10.1038/gt.2014.37.
PMID:
24830436
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24830436
Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines.
Walters AA1, Kinnear E1, Shattock RJ1, McDonald JU1, Caproni LJ2, Porter N2, Tregoning JS1.
Author information
Abstract
The use of DNA to deliver vaccine antigens offers many advantages, including ease of manufacture and cost. However, most DNA vaccines are plasmids and must be grown in bacterial culture, necessitating elements that are either unnecessary for effective gene delivery (for example, bacterial origins of replication) or undesirable (for example, antibiotic resistance genes). Removing these elements may improve the safety profile of DNA for the delivery of vaccines. Here, we describe a novel, double-stranded, linear DNA construct produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, polyA tail and telomeric ends. We compared these constructs (called 'Doggybones' because of their shape) with conventional plasmid DNA. Using luciferase-expressing constructs, we demonstrated that expression levels were equivalent between Doggybones and plasmids both in vitro and in vivo. When mice were immunized with DNA constructs expressing the HIV envelope protein gp140, equivalent humoral and cellular responses were induced. Immunizations with either construct type expressing hemagluttinin were protective against H1N1 influenza challenge. This is the first example of an effective DNA vaccine, which can be produced on a large scale by enzymatic processes.Gene Therapy advance online publication, 15 May 2014; doi:10.1038/gt.2014.37.
PMID:
24830436
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24830436