Citrus tristeza virus based vectors for foreign gene/S expression
Disclosed herein are viral vectors based on modifications of the Citrus Tristeza virus useful for transfecting citrus trees for beneficial purposes. Included in the disclosure are viral vectors including one or more gene cassettes that encode heterologous polypeptides. The gene cassettes are positioned at desirable locations on the viral genome so as to enable expression while preserving functionality of the virus. Also disclosed are methods of transfecting plants and plants transfected with viral vector embodiments.
1. A method of transfecting a citrus tree with a gene of interest having a 5′ end and a 3′ end, said method comprising inoculating said citrus tree with a sample comprising at least one Citrus tristeza virus (CTV) vector to produce an inoculated tree, said CTV vector engineered to comprise a construct comprising a heterologous gene of interest and a subgenomic RNA (sgRNA) heterologous controller element (CE) positioned upstream of the 5′ end of said heterologous gene of interest so as to control expression of said heterologous gene of interest; and growing said inoculated tree under conditions to allow a systemic infection of said tree with said at least one CTV vector; wherein expression of said heterologous gene of interest occurs for two months or more in the tree with a majority of virus in the systemic infection comprising said CTV vector.