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Targeted RNA recombination for the mutagenesis of coronavirus genomes

Reference	BBS/E/I/00000704
Principal Investigator / Supervisor	Professor Paul Britton
Co-Investigators / Co-Supervisors
Institution	The Pirbright Institute
Department	The Pirbright Institute Department
Funding type	Research
Value (£)	116,220
Status	Completed
Type	Institute Project
Start date	01/10/1998
End date	30/09/2001
Duration	36 months

Abstract

Recent work involving the coronavirus mouse hepatitis virus (MHV) has demonstrated that targeted recombination, using replicating defective RNAs (D-RNA), is a plausible way of introducing specific mutations into the virus genome. Our recent work has involved the isolation and characterisation of a D-RNA, derived from the avian coronavirus infectious bronchitis virus (IBV), that can be generated from a cDNA copy and rescued by helper virus following serial passage. We have demonstrated that reporter genes can be expressed from within the IBV replicating D-RNA. We have also demonstrated that the Beaudette derived D-RNA can be rescued by other IBV strains and that the leader sequences of the rescued D- RNAs are derived from the helper strain. This observation indicates a recombination event between the input D-RNA and the helper virus genome. The aim of this project is to utilise the D- RNA as a method for targeted recombination in IBV. The project will also investigate the rescue of IBV D-RNAs in ovo to determine whether recombination is more feasible in an in vivo system rather than using cell culture. The IBV D-RNA we are presently using was derived from the Beaudeete strain of IBV and although we have rescued this D-RNA with several other strains of IBV the efficiency of rescue varies and that the leader sequence on the rescued D-RNAs is exchanged for that of the helper virus. The project will investigate whether exchanging the 3that and 5that untranslated regions (UTRs) of the D-RNAs with those of the helper strain will improve the efficiency of rescue.

Summary

unavailable

Committee	Closed Committee - Genes & Developmental Biology (GDB)
Research Topics	X – not assigned to a current Research Topic
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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