Compositions and Methods for Targeted Delivery of CRISPR-CAS Effector Polypeptides and Transgenes
AssigneeThe Regents of the University of California
InventorsJennifer A. Doudna, Jennifer Hamilton, Connor Tsuchida
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Abstract
The present disclosure provides virus-like particles (VLPs) comprising: i) a CRISPR-Cas effector polypeptide; ii) a recombinant lentivirus comprising a nucleotide sequence encoding a therapeutic polypeptide having a length of from about 250 amino acids to about 3,000 amino acids, where the VLP comprises a pseudotyping viral glycoprotein and/or a polypeptide that provides for binding to a target cell. The present disclosure provides systems for producing a VLP. The present disclosure provides methods of delivering a therapeutic protein, using a VLP of the present disclosure.
Claims37
A system comprising:
- a) a first nucleic acid comprising a nucleotide sequence encoding a fusion polypeptide comprising:
- i) a lentiviral gag polyprotein comprising a matrix (MA) polypeptide, a capsid (CA) polypeptide, and a nucleocapsid (NC) polypeptide;
- and
- ii) a CRISPR-Cas effector polypeptide;
- wherein the fusion polypeptide comprises proteolytically cleavable linker between the gag polyprotein and the CRISPR-Cas effector polypeptide;
- b) a second nucleic acid comprising a nucleotide sequence encoding a therapeutic polypeptide, wherein the second nucleic acid comprises a recombinant lentiviral nucleic acid;
- c) a third nucleic acid comprising a nucleotide sequence encoding a pseudotyping viral envelope protein and/or a polypeptide that provides for binding to a target cell;
- and
- d) a fourth nucleic acid comprising a nucleotide sequence encoding a lentiviral pol polyprotein comprising a reverse transcriptase and an integrase;
- wherein the system comprises a nucleic acid comprising a nucleotide sequence encoding a CRISPR-Cas guide RNA, wherein the CRISPR-Cas guide RNA-encoding nucleic acid is either:
- i) part of the second nucleic acid;
- or
- ii) a fifth nucleic acid.
A virus-like particle (VLP) comprising:
- a) a CRISPR-Cas effector polypeptide;
- and
- b) a recombinant lentivirus comprising a nucleotide sequence encoding a therapeutic polypeptide having a length of from about 250 amino acids to about 3,000 amino acids,
- wherein the VLP comprises a pseudotyping viral glycoprotein and/or a polypeptide that provides for binding to a target cell.
Dependencies are inferred from the claim text.