Retroviral particle comprising at least two encapsidated nonviral RNAs
The present invention relates to a retroviral system for the transfer of non-viral RNA into target cells and more particularly a retroviral particle capable of delivering multiple RNAs. More particularly, it relates to retroviral particles comprising a protein derived from the Gag polyprotein an envelope protein, optionally an integrase and at least two encapsidated non-viral RNAs, the encapsidated non-viral RNAs each comprising an RNA sequence of interest linked to an encapsidation sequence, each encapsidation sequence being recognised by a binding domain introduced into the protein derived from the Gag polyprotein and/or into the integrase.
1. A retroviral particle comprising a protein from a Gag polyprotein, an envelope protein, optionally an integrase, and at least two encapsidated, different, non-viral RNAs, the at least two encapsidated, different, non-viral RNAs each comprising an RNA sequence of interest linked to an encapsidation sequence, each encapsidation sequence bound to a heterologous binding domain introduced into the protein from the Gag polyprotein.
2. The retroviral particle according to claim 1 , wherein the protein from the Gag polyprotein is a nucleocapsid protein.
3. The retroviral particle according to claim 2 , which is a lentiviral particle.
4. The retroviral particle which is a lentiviral particle according to claim 3 , wherein:
the binding domain is the Coat protein of bacteriophage MS2;
each of the encapsidation sequences of the non-viral RNAs is a stem-loop sequence of MS2; and
the nucleocapsid protein is the nucleocapsid protein (NC) of HIV, wherein the Coat protein is inserted at the second zinc finger of the NC.
5. The retroviral particle which is a lentiviral particle according to claim 3 , wherein:
the binding domain is the Coat protein of bacteriophage PP7;
each of the encapsidation sequences of the non-viral RNAs is a stem-loop sequence of PP7; and
the nucleocapsid protein is the nucleocapsid protein (NC) of HIV, wherein the Coat protein is inserted at the second zinc finger of the NC.
6. A composition comprising a plurality of the retroviral particle according to claim 1 .