ENGINEERED HUMAN-ENDOGENOUS VIRUS-LIKE PARTICLES AND METHODS OF USE THEREOF FOR DELIVERY TO CELLS

1 . An engineered human-derived virus-like particle (heVLP), comprising: a membrane comprising a phospholipid bilayer with one or more HERV-derived ENV/glycoprotein(s) on the external side; a HERV-derived GAG protein in the heVLP core, and a cargo disposed in the core of the heVLP on the inside of the membrane, wherein the cargo is fused to a human-endogenous GAG or other plasma membrane recruitment domain, and the heVLP does not comprise a non-human gag and/or pol protein.

2 . The heVLP of claim 1 , wherein the cargo is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein, or a small molecule.

3 . The heVLP of claim 1 , wherein the cargo is a gene editing reagent.

4 . The heVLP of claim 1 , wherein the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.

5 . The heVLP of claim 4 , wherein the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.

6 . The heVLP of claim 4 , wherein the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.

7 . The heVLP of claim 1 , wherein the cargo comprises a fusion to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6.

8 . A method of delivering a cargo molecule to a target cell, optionally a cell in vivo or in vitro, the method comprising contacting the cell with the heVLP of claim 1 comprising the cargo molecule, preferably wherein the cargo molecule is a biomolecule and/or chemical.

9 . A method of producing a heVLP comprising one or more cargo molecules, the method comprising:

providing a cell expressing one or more HERV-derived envelope proteins, one or more HERV-derived GAG proteins, and the one or more cargo molecules, wherein the cell does not express a gag and/or pol protein, except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome; and

maintaining the cell under conditions such that the cells produce heVLPs.

10 . The method of claim 9 , further comprising harvesting and optionally purifying and/or concentrating the produced heVLPs.

11 . The method of claim 9 , wherein the cargo molecule is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein, or a small molecule therapeutic or diagnostic agent.

12 . The method of claim 9 , wherein the cargo molecule is a gene editing reagent.

13 . The method of claim 9 , wherein the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.

14 . The method of claim 13 , wherein the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.

15 . The method of claim 13 , wherein the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.

16 . The method of claim 9 , wherein the cargo molecule comprises a fusion to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6.

17 . A cell that expresses, in combination:

one or more HERV-derived envelope proteins;

one or more HERV-derived GAG proteins; and

a cargo molecule, preferably fused to a human endogenous GAG or other plasma membrane recruitment domain,

wherein the cell does not express non-human gag protein.

18 . The cell of claim 17 , wherein the cargo molecule is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein.

19 . The cell of claim 17 , wherein the cargo molecule is a gene editing reagent.

20 . The cell of claim 17 , wherein the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.

21 . The cell of claim 20 , wherein the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.

22 . The cell of claim 20 , wherein the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.

23 . The cells of claim 17 , wherein the cargo molecule comprises a fusion to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6.

24 . The cells of claim 17 , wherein the cells are primary or stable human cell lines.

25 . The cells of claim 24 , which are Human Embryonic Kidney (HEK) 293 cells, HEK293 T cells, or BeWo cells.