Kits comprising plus-sense single stranded RNA viral vectors and methods for producing polypeptides using the kits

A mixture comprising:

• a) at least one first plus-sense single stranded RNA viral vector,
• b) at least one second plus-sense single stranded RNA viral vector, and, optionally,
• c) at least one third plus-sense single stranded RNA viral vector,
• wherein
• (i) the first plus-sense single-stranded viral vector and the second plus-sense single-stranded viral vector are derived from different plant viruses, and
• (ii) the coat protein ORF of the virus from which the first vector is derived is completely deleted in the first plus-sense single stranded RNA viral vector, and
• (iii) the coat protein ORF of the virus from which the second vector is derived is completely deleted in the second plus-sense single stranded RNA viral vector, and
• (iv) the first plus-sense single stranded RNA viral vector comprises a functional coat protein ORF of the virus from which the second plus-sense single-stranded viral vector is derived, and
• (v) the second plus-sense single stranded RNA viral vector comprises a functional coat protein ORF of the virus from which the first plus-sense single-stranded viral vector is derived, and
• (vi) the first plus-sense single-stranded viral vector and the second plus-sense single-stranded viral vector comprise an RNA replicon which is able to replicate in plant cells, and
• wherein
• the virus from which the first plus-sense single-stranded viral vector is derived is a Potexvirus,
• the virus from which the second plus-sense single-stranded viral vector is derived is a Tobamovirus, and
• the at least one third plus-sense single-stranded viral vector is derived from a Potexvirus.

A plant or plant cell

• comprising at least one first plus-sense single stranded RNA viral vector and at least one second plus-sense single stranded RNA viral vector, and, optionally, at least one third plus-sense single stranded RNA viral vector(s),
• wherein
• (i)the first plus-sense single-stranded viral vector and the second plus-sense single-stranded viral vector are derived from different plant viruses, and
• (ii) the coat protein ORF of the virus from which the first vector is derived is completely deleted in the first plus-sense single stranded RNA viral vector, and
• (iii) the coat protein ORF of the virus from which the second vector is derived is completely deleted in the second plus-sense single stranded RNA viral vector, and
• (iv) the first plus-sense single stranded RNA viral vector comprises a functional coat protein ORF of the virus from which the second plus-sense single-stranded viral vector is derived, and
• (v) the second plus-sense single stranded RNA viral vector comprises a functional coat protein ORF of the virus from which the first plus-sense single-stranded viral vector is derived, and
• (vi) the first plus-sense single-stranded viral vector and the second plus-sense single-stranded viral vector comprise an RNA replicon which is able to replicate in plant cells, and
• wherein
• the virus from which the first plus-sense single-stranded viral vector is derived is a Potexvirus, and
• the virus from which the second plus-sense single-stranded viral vector is derived is a Tobamovirus, and
• the at least one third plus-sense single-stranded viral vector is derived from a Potexvirus.

A method for producing in a plant, or-plant tissue, or plant cell a heterooligomeric polypeptide and/or two or more polypeptides, comprising providing to at least one plant cell

• a)at least one first plus-sense single stranded RNA viral vector, and
• b)at least one second plus-sense single stranded RNA viral vector, and
• c)optionally, at least one third plus-sense single stranded RNA viral vector,
• wherein
• (i)the first plus-sense single-stranded viral vector and the second plus-sense single-stranded viral vector are derived from different plant viruses, and
• (ii) the coat protein ORF of the virus from which the first vector is derived is completely deleted in the first plus-sense single stranded RNA viral vector, and
• (iii) the coat protein ORF of the virus from which the second vector is derived is completely deleted in the second plus-sense single stranded RNA viral vector, and
• (iv) the first plus-sense single stranded RNA viral vector comprises a functional coat protein ORF of the virus from which the second plus-sense single-stranded viral vector is derived, and
• (v) the second plus-sense single stranded RNA viral vector comprises a functional coat protein ORF of the virus from which the first plus-sense single-stranded viral vector is derived, and
• (vi) the first plus-sense single-stranded viral vector and the second plus-sense single-stranded viral vector comprise an RNA replicon which is able to replicate in plant cells, and
• (vii) the at least one plus-sense single stranded RNA viral vector and the at least one second plus-sense single stranded RNA viral vector comprise different heterologous ORFs, and
• wherein
• the virus from which the first plus-sense single-stranded viral vector is derived is a Potexvirus, and
• the virus from which the second plus-sense single-stranded viral vector is derived is a Tobamovirus, and
• the at least one third plus-sense single-stranded viral vector is derived from a Potexvirus.