UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Construction and characterisation of models for X-linked severe combined immunodeficiency for targeted gene correction by zinc finger nucleases.

Pallant, CE; (2013) Construction and characterisation of models for X-linked severe combined immunodeficiency for targeted gene correction by zinc finger nucleases. Doctoral thesis , UCL.

Full text not available from this repository.

Abstract

X-linked Severe Combined Immunodeficiency (SCID-X1) is an immunopathy caused by a mutation of the common gamma chain (γc) gene, IL2RG, which results in a lack of T cells, NK cells and with dysfunctional B cells. Current gene therapy methods involve the addition of a correct γc gene via integrating viral vectors. However, these current non-targeting gene addition strategies can result in transformation of the cell. A novel solution to this problem is met by targeted gene correction via homologous recombination stimulated by a site specific cleavage event caused by zinc finger nucleases (ZFN) within the disease gene. A γc deficient mouse has been created by replacing the murine il2rg locus with a mutated human IL2RG containing a point mutation frequently seen in SCID-X1 patients. The mutant human IL2RG is transcribed and initial analysis of this new SCID-X1 model has revealed a phenotype mirroring γc gene knockout mice. Lineage negative bone marrow cells from these mice, transduced with integrating lentiviral vector encoding functional IL2RG can reconstitute the immune cells in the Rag2-/-γc-/- double knockout SCID mouse model. Therefore the humanised mouse model of SCID-X1 can be corrected and is an appropriate platform to assess the efficiency of various gene targeting and correction strategies for the human mutation including ZFN induced homologous recombination. We have successfully achieved targeted homologous recombination in both a human T cell SCID-X1 cell line model and the humanised mouse embryonic stem cells with IL2RG specific ZFN.

Type: Thesis (Doctoral)
Title: Construction and characterisation of models for X-linked severe combined immunodeficiency for targeted gene correction by zinc finger nucleases.
Event: UCL
URI: https://discovery.ucl.ac.uk/id/eprint/1403521
Downloads since deposit
0Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item