Design: Genetic elements
Key objectives
Small circular episomes, constructed from defined genetic components, hold great promise for various applications in applied sciences and biotechnology, including gene therapy. Therapeutic application demands that the genetic elements, which are required to regulate fundamental aspects of vector function, can be defined precisely. In this section prototype vectors with defined reporter gene expression cassettes will be used to establish the genetic elements that determine optimal vector performance. The key objectives will focus on defining: The role of genetic elements in defining the mechanisms by which replication of episomal DNA is regulated and the molecular mechanisms that dictate the efficiency of this process.
- The importance of S/MAR structure and genetic configuration in defining the extra-chromosomal behavior and optimal performance of episomal gene expression systems.
- The critical features of gene architecture that will deliver regulated gene expression from mini-gene constructs expressed on episomal gene delivery vectors in different cell types.
- Protocols for the development of gene delivery vectors that are derived only from precisely defined genetic elements of human origin.
Deliverables
The part of the project will develop prototypes of circular vectors to provide refined vectors. Vectors will be developed that contain reporter genes or therapeutic genes with tissue specific control elements as well as vectors with chromosomal origins of replication, and vectors with increased cloning capacity.