Creating knockout tissues
Dr Paul Genever & Dr Gary Spencer
University of York
2007 – 2010 Research Associate
Generation of three-dimensional human tissue structures with targeted gene disruptions.
Dr Paul Genever is Head of the Biomedical Tissue Research Group in the Department of Biology at York University
Dr Gary Spencer is a Research Associate in the Department of Biology, York University.
Although the human genome has been sequenced, the function of each gene is still not known. A common approach to understanding gene function is to knock out genes in animals to see what, if any, effect can be observed.
Targeted gene disruption, usually in mice, is frequently used to determine the function of a particular gene or genes, often with the intention of understanding the genetic basis of human disease. The generation of knockout animals is technically demanding, time consuming and has low efficiency. Significantly, it is unclear how accurately the data obtained from knockout mice translates to human physiology.
There is a growing trend in the use of genetically modified animals, and the generation and maintenance of single knockout mouse ‘models’ can depend on extensive breeding programmes that require large numbers of animals. The technology in this project has the potential to replace and reduce animals in gene knockout experiments.
Mesenchymal stem cells (MSCs) are rare, multipotent cells that can be extracted from adult tissues, such as bone marrow. MSCs can be induced to undergo differentiation into a range of functional cell and tissue types, including bone, cartilage, fat and possibly other non-mesenchymal tissue including liver, brain and cardiac muscle. There is considerable interest in determining how MSCs may be used as in vitro models for fundamental research and drug discovery.
This Dr Hadwen Trust-funded project will use novel models of human MSC growth to generate 3D tissue structures with targeted gene disruptions, for functional analysis. An initial target will be a gene encoding a well-characterised transcription factor involved in MSC differentiation (Cbfa-1), which will have clearly defined measurable end-points. Also targeted will be relevant Wnt genes, as key regulators of mesenchymal differentiation. This work will, for the first time, identify the potential for using MSCs to generate “knockout tissues” with pervasive applications in biotechnology and basic research.