Viral guide rna delivery
OwnerFei Ran, ChieYu Lininventors
Life of this patent
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Abstract
Provided herein are recombinant negative-strand RNA virus genomes (e.g., recombinant rabies virus genomes) and recombinant negative-strand RNA viruses (e.g., recombinant rabies viruses) and methods for their use in delivering a guide RNA and, optionally, a transgene, into a target cell. Also provided are packaging systems and methods of using the packaging systems to produce recombinant negative-strand RNA viruses.
Claims39
A recombinant negative-strand RNA virus genome, comprising
- a nucleic acid encoding a first guide RNA (gRNA) that comprises a 5′ end and a 3′ end;
- and
- a nucleic acid encoding a first transfer RNA (tRNA) positioned at one or both of the 3′ end of the nucleic acid encoding the first gRNA or the 5′ end of the nucleic acid encoding the first gRNA.
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A recombinant rabies virus particle, comprising:
- a rabies virus glycoprotein;
- and
- a recombinant rabies virus genome comprising a nucleic acid encoding a first guide RNA (gRNA) that comprises a 5′ end and a 3′ end, and
- a nucleic acid encoding a first transfer RNA (tRNA) positioned at one or both of the 3′ end of the nucleic acid encoding the first gRNA or the 5′ end of the nucleic acid encoding the first gRNA.
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A method for expressing a nucleobase editor and guide RNA (gRNA) in a target cell, comprising transducing a target cell with a recombinant rabies virus particle, wherein the recombinant virus particle comprises:
- a rabies virus glycoprotein;
- and
- a recombinant rabies virus genome comprising:
- a nucleic acid encoding a nucleobase editor comprising a polynucleotide programmable nucleotide binding domain and a nucleobase editing domain;
- a nucleic acid encoding a first gRNA that comprises a 5′ end and a 3′ end;
- and
- a nucleic acid encoding a first tRNA positioned at one or both of the 3′ end of the nucleic acid encoding the first gRNA or the 5′ end of the nucleic acid encoding the first gRNA.
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A packaging system for the recombinant preparation of a rabies virus particle, wherein the packaging system comprises:
- an N gene encoding for a rabies virus nucleoprotein or a functional variant thereof;
- a P gene encoding for a rabies virus phosphoprotein or a functional variant thereof;
- an L gene encoding for a rabies virus polymerase or a functional variant thereof;
- and
- a recombinant rabies virus genome, wherein:
- the genome comprises a nucleic acid encoding a first guide RNA (gRNA) that comprises a 5′ end and a 3′ end;
- and
- the genome comprises a nucleic acid encoding a first transfer RNA (tRNA) positioned at one or both of the 3′ end of the nucleic acid encoding the first gRNA or the 5′ end of the nucleic acid encoding the first gRNA.
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Dependencies are inferred from the claim text.