Award details

Recombinase-mediated site-specific gene integration in mammalian somatic cells.

Reference	BB/C003543/1
Principal Investigator / Supervisor	Professor Marshall Stark
Co-Investigators / Co-Supervisors
Institution	University of Glasgow
Department	Institute of Biomedical & Life Sciences
Funding type	Research
Value (£)	235,856
Status	Completed
Type	Research Grant
Start date	01/03/2005
End date	31/07/2008
Duration	41 months

Abstract

Transgenes, introduced for therapeutic, biotechnology, or basic research purposes, should ideally be integrated at specific genomic loci, in order to ensure their safe and efficient expression. We have recently shown that chimaeric site-specific recombinases (Z-resolvases), based on the bacterial Tn3 resolvase, promote recombination at designed novel DNA sites. We now intend to develop these Z-resolvases further, to target chosen natural genomic sites in vivo. Creation of a Z-resolvase to target a natural site will require selection of DNA-binding domains (derived from Zif268) with specificity for the site sequences, using the phage display method, and optimisation of the recombinase catalytic domain by mutagenesis-selection. In this project, we aim to demonstrate the insertion of single copies of foreign genes into predefined positions in the genomes of mammalian cells; specifically, at sites within the bovine beta-casein gene, to facilitate the efficient production of pharmaceutically relevant proteins in milk. Z-resolvase targeted to a site in the casein gene will be introduced into, or expressed in the bovine cells, in the presence of transfected foreign gene DNA in a form appropriate for recombinase activity. The efficiency of targeted integration of the transgene will then be assessed by various methods. This project will also serve as a pilot study to evaluate the more general potential of these enzymes for promoting targeted recombination, for example in gene therapy.

Summary

unavailable

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
Research Topics	Pharmaceuticals, Technology and Methods Development
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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