Targeted lipid particles and compositions and their use

Abstract

Provided herein are lipid particles containing a lipid bilayer surrounding a lumen or cavity, a henipavirus F protein molecule or a biologically active portion thereof, and a targeting envelope protein containing a henipavirus envelope-associated glycoprotein G (G protein) or a biologically active portion thereof and a binding domain, e.g., a single-domain antibody (sdAb) variable domain. Also provided herein are targeting envelope proteins containing a G protein fused or linked to a binding domain, e.g., an sdAb variable domain, and polynucleotides encoding such proteins. Also provided are producer cells and compositions containing such targeted lipid particles, as well as methods for making and using targeted lipid particles.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of U.S. Provisional Application No. 63 / 003,168, filed Mar. 31, 2020, entitled “Targeted Lipid Particles and Compositions and Uses Thereof,” and U.S. Provisional Application No. 63 / 154,341, filed Feb. 26, 2021, entitled “Targeted Lipid Particles and Compositions and Uses Thereof,” the contents of each of which are hereby incorporated by reference in their entirety for all purposes.

[0002] Incorporation by Reference of Sequence Listing This application has been filed with an electronic sequence listing. The sequence listing is provided as a file entitled 186152003640SeqList.TXT, created on Mar. 29, 2021, with a size of 2,071,683 bytes. The electronic form information of the sequence listing is hereby incorporated by reference in its entirety.

[0003] Field The present disclosure relates to lipid particles containing a lipid bilayer surrounding a lumen or cavity, a henipavirus F protein molecule or a biologically active portion thereof, and a targeted envelope protein containing a henipavirus envelope - binding glycoprotein G (G protein) or a biologically active portion and binding domain thereof, such as a single - domain antibody (sdAb) variable domain. The present disclosure also provides a targeted envelope protein containing a G protein fused or linked to a binding domain, such as an sdAb variable domain, and a polynucleotide encoding such a protein. Also disclosed are producer cells and compositions containing such targeted lipid particles and methods of making and using the targeted lipid particles.

Background Art

[0004] Background Lipid particles, including virus-like particles and viral vectors, are commonly used for the delivery of exogenous substances to cells. However, the delivery of lipid particles to designated target cells can present challenges. In the case of lentiviral vectors, the host range can be altered by pseudotyped with heterologous envelope proteins. The designated retargeting envelope protein may not be sufficiently stable or even expressed on the surface of the lipid particle. Improved lipid particles, including virus-like particles and viral vectors, are needed to target desired cells. The provided disclosure addresses this need. [Overview of the Initiative]

[0005] overview This specification provides targeted lipid particles comprising (a) a lipid bilayer surrounding a lumen, (b) a henipavirus F protein molecule or a biologically active moiety thereof, and (c) a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof and (ii) a single-domain antibody (sdAb) variable domain, wherein the sdAb variable domain is bound to the C-terminus of the G protein or a biologically active moiety thereof, and the F protein molecule or a biologically active moiety thereof and the targeted envelope protein are embedded in the lipid bilayer. In some embodiments, the single-domain antibody is bound to the G protein via a linker. In some embodiments, the linker is a peptide linker.

[0006] This specification provides targeted lipid particles comprising (a) a lipid bilayer surrounding a lumen, (b) a henipavirus F protein molecule or its biologically active moiety, and (c) a targeted envelope protein comprising a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety, conjugated via a peptide linker to a variable domain of a single-domain antibody (sdAb), wherein the single-domain antibody is conjugated to a cell surface molecule of a target cell, and the F protein molecule or its biologically active moiety and the targeted envelope protein are embedded in the lipid bilayer. In some embodiments, the N-terminus of the F protein molecule or its biologically active moiety is exposed to the outside of the lipid bilayer. In some embodiments, the C-terminus of the G protein is exposed to the outside of the lipid bilayer.

[0007] In some embodiments, the single-domain antibody binds to a cell surface molecule present on the target cell. In some embodiments, the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule. In some of the optional embodiments, the single-domain antibody binds to an antigen or a portion thereof present on the target cell. In some embodiments, the antigen is a cell surface molecule or a portion thereof containing an epitope recognized by the single-domain antibody. In some of the optional embodiments, the target cell is selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. In some embodiments, the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytic cells, SLC7A10+ adipocytes, or CD30+ lung epithelial cells. In some of the embodiments, the target cells are hepatocytes. In some of the embodiments, the cell surface molecules or antigens are selected from the group consisting of ASGR1, ASGR2, and TM4SF5.

[0008] In some of the optional embodiments, the target cells are T cells. In some of the optional embodiments, the cell surface molecule or antigen is CD8 or CD4.

[0009] In some of the various embodiments, the cell surface molecule or antigen is LDL-R.

[0010] This specification provides a targeted lipid particle comprising (a) a lipid bilayer surrounding the lumen, (b) a henipavirus F protein molecule or its biologically active portion, and (c) a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active portion, the binding domain binds to a cell surface molecule selected from the group consisting of ASGR1, ASGR2, and TM4SF5, and optionally human ASGR1, human ASGR2, and human ASGR2, and the F protein molecule or its biologically active portion and the targeted envelope protein are embedded in the lipid bilayer.

[0011] This specification provides a targeted lipid particle comprising (a) a lipid bilayer surrounding a lumen, (b) a henipavirus F protein molecule or its biologically active portion, and (c) a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active portion, the binding domain binds to a cell surface molecule selected from the group consisting of CD8 and CD4, optionally human CD8 or human CD4, and the F protein molecule or its biologically active portion and the targeted envelope protein are embedded in the lipid bilayer.

[0012] This specification provides a targeted lipid particle comprising (a) a lipid bilayer surrounding a lumen, (b) a henipavirus F protein molecule or its biologically active portion, and (c) a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active portion, the binding domain binds to a cell surface molecule which is a low-density lipoprotein receptor (LDL-R), optionally human LDL-R, and the F protein molecule or its biologically active portion and the targeted envelope protein are embedded in the lipid bilayer.

[0013] In some of the optional embodiments, the lipid particles are lentiviral vectors. In some of the optional embodiments, the binding domain is linked to a G protein via a linker. In some of the optional embodiments, the linker is a peptide linker.

[0014] This specification provides a lentiviral vector comprising a binding domain that targets a cell surface molecule selected optionally from the group consisting of ASGR1, ASGR2, and TM4SF5, and is pseudotyped with a retargeting viral fusion protein, wherein the retargeting viral fusion protein comprises (a) a henipavirus F protein molecule or its biologically active portion, and (b) a targeted envelope protein comprising a binding domain bound to henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion.

[0015] This specification provides a lentiviral vector comprising a binding domain that targets cell surface molecules selected from the group consisting of CD8 and CD4, optionally human CD8 and human CD4, and which is pseudotyped with a retargeting viral fusion protein, wherein the retargeting viral fusion protein comprises (a) a henipavirus F protein molecule or its biologically active portion, and (b) a targeted envelope protein comprising a binding domain bound to henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion.

[0016] This specification provides a lentiviral vector comprising a binding domain that targets a low-density lipoprotein receptor (LDL-R), wherein the LDL-R is optionally human LDL-R, and the lentiviral vector is pseudotyped with a retargeting viral fusion protein comprising (a) a henipavirus F protein molecule or its biologically active portion, and (b) a targeted envelope protein comprising a binding domain bound to henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion.

[0017] In some of the available embodiments, the binding domain is bound to the C-terminus of the G protein or its biologically active portion.

[0018] In this specification, (a) Henipavirus F protein molecule or its biologically active portion, (b) A targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active moiety, and the binding domain binds to CD4, (c) A cargo comprising a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR is (i) an extracellular antigen-binding domain that binds to an extracellular antigen (e.g., CD19 or BCMA), and (ii) Intracellular signaling domains comprising a CD3 zeta signaling domain and optionally a 4-1BB or CD28 costimulatory signaling domain Including cargo and A lentiviral vector is provided that includes the following. In some embodiments, the extracellular antigen-binding domain of the CAR is scFv.

[0019] In some of the available embodiments, the lentiviral vector is capable of delivering CAR-encoding nucleic acids to T cells. In some embodiments, the T cells are in vivo in the subject.

[0020] This specification provides a lentiviral vector comprising (a) a henipavirus F protein molecule or a biologically active moiety thereof, and (b)(i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof, and (ii) a targeted envelope protein comprising a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or the biologically active moiety thereof, the binding domain binds to ASGR1, and the lentiviral vector is capable of targeting hepatocytes. In some of the various embodiments, the lentiviral vector further comprises an exogenous substance for delivery to hepatocytes.

[0021] In some of the optional embodiments, the lentiviral vector is capable of delivering exogenous material to hepatocytes, which are optionally in vivo in the subject.

[0022] In some of the embodiments, the binding domain is bound to a G protein via a linker. In some of the embodiments, the linker is a peptide linker. In some of the embodiments, the binding domain is a single-domain antibody. In some of the embodiments, the binding domain is a single-stranded variable fragment (scFv).

[0023] In some of the optional embodiments, the peptide linker has a length of up to 65 amino acids. In some of the optional embodiments, the peptide linker has a length of up to 50 amino acids. In some of the optional embodiments, the peptide linker has a length of approximately 2-65 amino acids, 2-60 amino acids, 2-56 amino acids, 2-52 amino acids, 2-48 amino acids, 2-44 amino acids, 2-40 amino acids, 2-36 amino acids, 2-32 amino acids, 2-28 amino acids, 2-24 amino acids, 2-20 amino acids, 2-18 amino acids, 2-14 amino acids, 2-12 amino acids, 2-10 amino acids, 2-8 amino acids, 2-6 amino acids, 6-65 amino acids, 6-60 amino acids, 6-56 amino acids, 6-52 amino acids, 6-48 amino acids Amino acids, 6-44 amino acids, 6-40 amino acids, 6-36 amino acids, 6-32 amino acids, 6-28 amino acids, 6-24 amino acids, 6-20 amino acids, 6-18 amino acids, 6-14 amino acids, 6-12 amino acids, 6-10 amino acids, 6-8 amino acids, 8-65 amino acids, 8-60 amino acids, 8-56 amino acids, 8-52 amino acids, 8-48 amino acids, 8-44 amino acids, 8-40 amino acids, 8-36 amino acids, 8-32 amino acids, 8-28 amino acids, 8-24 amino acids, 8-20 amino acids, 8-18 amino acids, 8-14 amino acids, 8-12 amino acids, 8-10 amino acids, 10-65 amino acids, 10-60 amino acids, 10-56 amino acids, 10-52 amino acids, 10-48 amino acids, 10-44 amino acids, 10-40 amino acids, 10-36 amino acids, 10-32 amino acids, 10-28 amino acids, 10-24 amino acids, 10-20 amino acids, 10-18 amino acids, 10-14 amino acids, 10-12 amino acids, 12-65 amino acids, 12-60 amino acids, 12-56 amino acids, 12-52 amino acids, 12-48 amino acids, 1 2-44 amino acids, 12-40 amino acids, 12-36 amino acids, 12-32 amino acids, 12-28 amino acids, 12-24 amino acids, 12-20 amino acids, 12-18 amino acids, 12-14 amino acids, 14-65 amino acids, 14-60 amino acids, 14-56 amino acids, 14-52 amino acids, 14-48 amino acids, 14-44 amino acids, 14-40 amino acids, 14-36 amino acids, 14-32 amino acids, 14-28 amino acids, 14-24 amino acids, 14-20 amino acids, 14-18 amino acids, 18-65 amino acids,18-60 amino acids, 18-56 amino acids, 18-52 amino acids, 18-48 amino acids, 18-44 amino acids, 18-40 amino acids, 18-36 amino acids, 18-32 amino acids, 18-28 amino acids, 18-24 amino acids, 18-20 amino acids, 20-65 amino acids, 20-60 amino acids, 20-56 amino acids, 20-52 amino acids, 20-48 amino acids, 20-44 amino acids, 20-40 amino acids, 20-36 amino acids, 20-32 amino acids, 20-28 amino acids, 20-2 6 amino acids, 20-24 amino acids, 24-65 amino acids, 24-60 amino acids, 24-56 amino acids, 24-52 amino acids, 24-48 amino acids, 24-44 amino acids, 24-40 amino acids, 24-36 amino acids, 24-32 amino acids, 24-30 amino acids, 24-28 amino acids, 28-65 amino acids, 28-60 amino acids, 28-56 amino acids, 28-52 amino acids, 28-48 amino acids, 28-44 amino acids, 28-40 amino acids, 28-36 amino acids, 28-34 amino acids Acid, 28-32 amino acids, 32-65 amino acids, 32-60 amino acids, 32-56 amino acids, 32-52 amino acids, 32-48 amino acids, 32-44 amino acids, 32-40 amino acids, 32-38 amino acids, 32-36 amino acids, 36-65 amino acids, 36-60 amino acids, 36-56 amino acids, 36-52 amino acids, 36-48 amino acids, 36-44 amino acids, 36-40 amino acids, 40-65 amino acids, 40-60 amino acids, 40-56 amino acids, 40-52 amino acids, 40 Contains ~48 amino acids, 40~44 amino acids, 44~65 amino acids, 44~60 amino acids, 44~56 amino acids, 44~52 amino acids, 44~48 amino acids, 48~65 amino acids, 48~60 amino acids, 48~56 amino acids, 48~52 amino acids, 50~65 amino acids, 50~60 amino acids, 50~56 amino acids, 50~52 amino acids, 54~65 amino acids, 54~60 amino acids, 54~56 amino acids, 58~65 amino acids, 58~60 amino acids, or 60~65 amino acids. In some of the optional embodiments, the peptide linker has lengths of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,The peptide contains polypeptides having 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65 amino acids. In some of the embodiments, the peptide linker is a flexible linker containing GS, GGS, GGGGS (SEQ ID NO: 43), GGGGGS (SEQ ID NO: 41), or a combination thereof. In some of the embodiments, the peptide linker contains (GGS)n (n is 1 to 10). In some of the embodiments, the peptide linker contains (GGGGS)n (SEQ ID NO: 42) (n is 1 to 10). In some of the embodiments, the peptide linker contains (GGGGGS)n (SEQ ID NO: 27) (n is 1 to 6).

[0024] In some of the optional embodiments, the G protein or its biologically active portion is the wild-type Nipah virus G (NiV-G) protein or the Hendra virus G protein. In some of the optional embodiments, the G protein or its biologically active portion is the wild-type NiV-G protein or a functionally active variant or biologically active portion thereof. In some of the embodiments, the mutant NiV-G protein or its functionally active variant or biologically active moiety contains an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44.

[0025] In some of the available embodiments, the NiV-G protein is a cleaved and biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that lacks up to 40 consecutive amino acid residues at or near its N-terminus.

[0026] In some of the arbitrary embodiments, the NiV-G protein is a biologically active portion cleaved at the N-terminus of wild-type NiV-G, and is a sequence shown in any of SEQ ID NOs: 10-15, 35-40 or 45-50, or a sequence with at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 8 It has an amino acid sequence having sequence identity of 6%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%.

[0027] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 5-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 10. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 35.In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 45.

[0028] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 10-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 36. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 11.In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 46.

[0029] In some of the optional embodiments, the NiV-G protein or the biologically active moiety has a 15-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44). In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 12. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 37.In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 47.

[0030] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 20-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 13. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 38.In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 48.

[0031] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 25-amino acid cleavage at or near its N-terminus. In some of the various embodiments, the NiV-G protein has an amino acid sequence that has at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 14. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 39.In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 49.

[0032] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 30-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 15. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 40.

[0033] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 34-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 22. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 53.

[0034] In some of the various embodiments, the G protein, its biologically active portion, is a functionally active variant, which is a mutant NiV-G protein exhibiting reduced binding to ephrin B2 or ephrin B3.

[0035] In some of the optional embodiments, the mutant NiV-G protein contains one or more amino acid substitutions selected from the group consisting of E501A, W504A, Q530A, and E533A, with reference to the numbering shown in Sequence ID No. 28.

[0036] In some of the embodiments, the mutant NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 16. In some of the embodiments, the mutant NiV-G protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 51.

[0037] In some of the optional embodiments, the F protein or its biologically active portion is the wild-type Nipah virus F (NiV-F) protein or the Hendra virus F protein, or a functionally active variant or biologically active portion thereof. In some of the embodiments, the NiV-F protein or its functionally active variant or biologically active moiety includes the amino acid sequence shown in SEQ ID NO: 2, or an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 2.

[0038] In some of the optional embodiments, the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 20-amino acid cleavage at or near its C-terminus.

[0039] In some of the embodiments, the NiV-F protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 5.

[0040] In some of the arbitrary embodiments, the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) that includes i) a 20-amino acid cleavage at or near the C-terminus, and ii) a point mutation at the N-linked glycosylation site.

[0041] In some of the embodiments, the NiV-F protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 7.

[0042] In some of the optional embodiments, the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 22-amino acid cleavage at or near its C-terminus.

[0043] In some of the various embodiments, the NiV-F protein or the biologically active portion has an amino acid sequence encoded by a sequence of nucleotides encoding the sequence shown in SEQ ID NO: 8, or a sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 8.

[0044] In some of the embodiments, the NiV-F protein or the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 23. In some of the various embodiments, the F-protein or its biologically active portion includes an F1 subunit or its fusion portion.

[0045] In some of the optional embodiments, the F protein comprises the sequence shown in SEQ ID NO: 23, and the G protein comprises the sequence shown in SEQ ID NO: 16.

[0046] In some of the various embodiments, the F protein consists of, or is essentially, the sequence shown in SEQ ID NO: 23, and / or the G protein consists of, or is essentially, the sequence shown in SEQ ID NO: 16.

[0047] In some of the various embodiments, the F1 subunit is a proteolytically cleaved portion of the F0 precursor. In some of the various embodiments, the F1 subunit has an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 4.

[0048] In some of the optional embodiments, the lipid bilayer is derived from the membrane of a host cell used to generate a retrovirus or retrovirus-like particle. In some of the optional embodiments, the host cell is selected from the group consisting of CHO cells, BHK cells, MDCK cells, C3H 10T1 / 2 cells, FLY cells, Psi-2 cells, BOSC 23 cells, PA317 cells, WEHI cells, COS cells, BSC 1 cells, BSC 40 cells, BMT 10 cells, VERO cells, W138 cells, MRC5 cells, A549 cells, HT1080 cells, 293 cells, 293T cells, B-50 cells, 3T3 cells, NIH3T3 cells, HepG2 cells, Saos-2 cells, Huh7 cells, HeLa cells, W163 cells, 211 cells, and 211A cells. In some of the optional embodiments, the host cell includes 293T cells. In some of the optional embodiments, the lipid bilayer is or contains a viral envelope. In some of the optional embodiments, the retrovirus-like particle is replication-deficient.

[0049] In some of the optional embodiments, the targeted lipid particles include one or more viral components other than F protein molecules and G proteins. In some of the optional embodiments, one or more viral components are from retroviruses. In some of the optional embodiments, the retrovirus is a lentivirus. In some of the optional embodiments, one or more viral components include a viral packaging protein selected from one or more of Gag, Pol, Rev, and Tat. In some of the optional embodiments, one or more viral components include one or more (e.g., all) of the following nucleic acid sequences: 5'LTR (e.g., containing U5 and lacking a functional U3 domain), psi packaging element (psi), central polypurine sequence (cPPT) / central termination sequence (CTS) (e.g., DNA flap), poly-A tail sequence, post-transcriptional regulatory element (e.g., WPRE), Rev reaction element (RRE), and 3'LTR (e.g., containing U5 and lacking a functional U3).

[0050] In some of the optional embodiments, the targeted lipid particles are lentiviral vectors.

[0051] In some of the optional embodiments, the targeted lipid particles or lentiviral vectors are replication-deficient.

[0052] In some of the optional embodiments, the targeted lipid particles or lentiviral vector further comprise exogenous material. In some of the optional embodiments, the targeted lipid particles further comprise exogenous material. In some embodiments, the lentiviral vector further comprises exogenous material.

[0053] In some of the optional embodiments, the exogenous substance is present in the lumen. In some of the optional embodiments, the exogenous substance is a protein or nucleic acid. In some embodiments, the nucleic acid is DNA or RNA.

[0054] In some of the optional embodiments, the exogenous substance is a nucleic acid encoding cargo for delivery to target cells. In some of the optional embodiments, the exogenous substance encodes a therapeutic or diagnostic agent.

[0055] In some of the various embodiments, the exogenous substance encodes a membrane protein. In some embodiments, the protein is an antigen receptor for targeting cells expressed by or associated with a disease or pathological condition. In some embodiments, the membrane protein is a chimeric antigen receptor (CAR). In some embodiments, the CAR comprises (i) an extracellular antigen-binding domain that binds to an extracellular antigen (e.g., CD19 or BCMA), optionally the extracellular antigen-binding domain being an scFv; (ii) a transmembrane domain; and (iii) an intracellular signaling region comprising a CD3 zeta signaling domain and optionally a co-stimulatory signaling domain, e.g., a 4-1BB or CD28 co-stimulatory signaling domain. In some embodiments, the target cell is a T cell. In some embodiments, the cell surface molecule on the target cell is CD4 or CD8. In some embodiments, the binding domain is an scFv that binds to CD4 (e.g., human CD4). In some embodiments, the binding domain is a single-domain antibody that binds to CD4 (e.g., human CD4). In some embodiments, the binding domain is an scFv that binds to CD8 (e.g., human CD8). In some embodiments, the binding domain is a single-domain antibody that binds to CD8 (e.g., human CD8).

[0056] In some of the optional embodiments, the exogenous substance is a nucleic acid containing a gene defect, optionally a payload gene for correcting the gene defect in target cells. In some embodiments, the gene defect is associated with liver cells or hepatocytes. In some embodiments, the target cells are hepatocytes. In some embodiments, the cell surface molecule is a molecule selected from the group consisting of ASGR1, ASGR2, and TM4SF5. In some embodiments, the binding domain is an scFv that binds to ASGR1 (e.g., human ASGR1). In some embodiments, the binding domain is a single-domain antibody that binds to ASGR1 (e.g., human ASGR1). In some embodiments, the binding domain is an scFv that binds to ASGR2 (e.g., human ASGR2). In some embodiments, the binding domain is a single-domain antibody that binds to ASGR2 (e.g., human ASGR2). In some embodiments, the binding domain is an scFv that binds to TM4SF5 (e.g., human TM4SF5). In some embodiments, the binding domain is a single-domain antibody that binds to TM4SF5 (e.g., human TM4SF5).

[0057] In some of the optional embodiments, the single-domain antibody binds to a cell surface molecule present on the target cell. In some of the optional embodiments, the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule. In some of the optional embodiments, the target cell is selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. In some of the optional embodiments, the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytocytes, SLC7A10+ adipocytes, or CD30+ lung epithelial cells.

[0058] In some of the optional embodiments, a single-domain antibody binds to an antigen or a portion thereof present on a target cell. In some of the optional embodiments, the cell surface molecule or antigen is selected from the group consisting of ASGR1, ASGR2, and TM4SF5. In some embodiments, the antigen or a portion thereof is human ASGR1. In some embodiments, the antigen or a portion thereof is human ASGR2. In some embodiments, the antigen or a portion thereof is human TM4SF5.

[0059] This specification provides polynucleotides comprising a nucleic acid sequence encoding (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof, and (ii) a binding domain that binds to a cell surface molecule selected from the group consisting of ASGR1, ASGR2, and TM4SF5. In some embodiments, the cell surface molecule is human ASGR1. In some embodiments, the cell surface molecule is human ASGR2. In some embodiments, the cell surface molecule is human TM4SF5. In some of the various embodiments, the cell surface molecule or antigen is CD8 or CD4.

[0060] This specification provides nucleic acid sequences encoding (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof, and (ii) a binding domain that binds to a cell surface molecule selected from the group consisting of CD4 and CD8. In some embodiments, the cell surface molecule is human CD4. In some embodiments, the cell surface molecule is human CD8. In some embodiments, the cell surface molecule or antigen is a low-density lipoprotein receptor (LDL-R). In some embodiments, the cell surface molecule or antigen is human LDL-R.

[0061] This specification provides polynucleotides comprising a nucleic acid sequence encoding (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and (ii) a binding domain that binds to a low-density lipoprotein receptor (LDL-R). In some embodiments, the binding domain binds to human LDL-R. In some of the embodiments, the binding domain is a single-domain antibody (sdAb). In some of the embodiments, the binding domain is a single-stranded variable fragment (scFv).

[0062] This specification provides a polynucleotide comprising a nucleic acid sequence encoding (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof and (ii) a single-domain antibody (sdAb) variable domain, wherein the sdAb variable domain is conjugated to the C-terminus of the G protein or the biologically active moiety thereof. In some of the various embodiments, the polynucleotide further comprises (iii) a nucleic acid sequence encoding a henipavirus F protein molecule or a biologically active moiety thereof.

[0063] In some embodiments, the nucleic acid sequence is a first nucleic acid sequence, and the polynucleotide further comprises a second nucleic acid sequence encoding the henipavirus F protein molecule or a biologically active portion thereof. In some embodiments, the polynucleotide comprises an IRES or a sequence encoding a linking peptide between the first and second nucleic acid sequences. In some embodiments, the linking peptide is a self-cleaving peptide or a peptide that causes ribosome skipping, optionally a T2A peptide.

[0064] In some of the optional embodiments, the polynucleotide includes at least one promoter functionally linked to control the expression of the nucleic acid. In some of the optional embodiments, the promoter is functionally linked to control the expression of a first nucleic acid sequence and a second nucleic acid sequence. In some of the optional embodiments, the promoter is a constitutive promoter. In some of the optional embodiments, the promoter is an inductive promoter.

[0065] In some of the optional embodiments, the sdAb variable domain is bound to the G protein via an encoded peptide linker. In some embodiments, the binding domain is bound to the G protein via an encoded peptide linker. In some of the optional embodiments, the encoded peptide linker has a length of up to 25 amino acids. In some of the optional embodiments, the encoded peptide linker has a length of up to 65 amino acids. In some of the optional embodiments, the encoded peptide linker has a length of approximately 2-65 amino acids, 2-60 amino acids, 2-56 amino acids, 2-52 amino acids, 2-48 amino acids, 2-44 amino acids, 2-40 amino acids, 2-36 amino acids, 2-32 amino acids, 2-28 amino acids, 2-24 amino acids, 2-20 amino acids, 2-18 amino acids, 2-14 amino acids, 2-12 amino acids, 2-10 amino acids, 2-8 amino acids, 2-6 amino acids, 6-65 amino acids. Amino acids, 6-60 amino acids, 6-56 amino acids, 6-52 amino acids, 6-48 amino acids, 6-44 amino acids, 6-40 amino acids, 6-36 amino acids, 6-32 amino acids, 6-28 amino acids, 6-24 amino acids, 6-20 amino acids, 6-18 amino acids, 6-14 amino acids, 6-12 amino acids, 6-10 amino acids, 6-8 amino acids, 8-65 amino acids, 8-60 amino acids, 8-56 amino acids, 8-52 amino acids, 8-48 amino acids, 8-44 Amino acids, 8-40 amino acids, 8-36 amino acids, 8-32 amino acids, 8-28 amino acids, 8-24 amino acids, 8-20 amino acids, 8-18 amino acids, 8-14 amino acids, 8-12 amino acids, 8-10 amino acids, 10-65 amino acids, 10-60 amino acids, 10-56 amino acids, 10-52 amino acids, 10-48 amino acids, 10-44 amino acids, 10-40 amino acids, 10-36 amino acids, 10-32 amino acids, 10-28 amino acids, 10 ~24 amino acids, 10~20 amino acids, 10~18 amino acids, 10~14 amino acids, 10~12 amino acids, 12~65 amino acids, 12~60 amino acids, 12~56 amino acids, 12~52 amino acids, 12~48 amino acids, 12~44 amino acids, 12~40 amino acids, 12~36 amino acids, 12~32 amino acids, 12~28 amino acids, 12~24 amino acids, 12~20 amino acids, 12~18 amino acids, 12~14 amino acids, 14~65 amino acids,14-60 amino acids, 14-56 amino acids, 14-52 amino acids, 14-48 amino acids, 14-44 amino acids, 14-40 amino acids, 14-36 amino acids, 14-32 amino acids, 14-28 amino acids, 14-24 amino acids, 14-20 amino acids, 14-18 amino acids, 18-65 amino acids, 18-60 amino acids, 18-56 amino acids, 18-52 amino acids, 18-48 amino acids, 18-44 amino acids, 18-40 amino acids, 18-36 amino acids, 18-32 amino acids, 18-28 amino acids, 18-24 amino acids, 18-20 amino acids, 20-65 amino acids Amino acids, 20-60 amino acids, 20-56 amino acids, 20-52 amino acids, 20-48 amino acids, 20-44 amino acids, 20-40 amino acids, 20-36 amino acids, 20-32 amino acids, 20-28 amino acids, 20-26 amino acids, 20-24 amino acids, 24-65 amino acids, 24-60 amino acids, 24-56 amino acids, 24-52 amino acids, 24-48 amino acids, 24-44 amino acids, 24-40 amino acids, 24-36 amino acids, 24-32 amino acids, 24-30 amino acids, 24-28 amino acids, 28-65 amino acids, 28-60 amino acids, 28- 56 amino acids, 28-52 amino acids, 28-48 amino acids, 28-44 amino acids, 28-40 amino acids, 28-36 amino acids, 28-34 amino acids, 28-32 amino acids, 32-65 amino acids, 32-60 amino acids, 32-56 amino acids, 32-52 amino acids, 32-48 amino acids, 32-44 amino acids, 32-40 amino acids, 32-38 amino acids, 32-36 amino acids, 36-65 amino acids, 36-60 amino acids, 36-56 amino acids, 36-52 amino acids, 36-48 amino acids, 36-44 amino acids, 36-40 amino acids, 40-65 amino acids It contains 40-60 amino acids, 40-56 amino acids, 40-52 amino acids, 40-48 amino acids, 40-44 amino acids, 44-65 amino acids, 44-60 amino acids, 44-56 amino acids, 44-52 amino acids, 44-48 amino acids, 48-65 amino acids, 48-60 amino acids, 48-56 amino acids, 48-52 amino acids, 50-65 amino acids, 50-60 amino acids, 50-56 amino acids, 50-52 amino acids, 54-65 amino acids, 54-60 amino acids, 54-56 amino acids, 58-65 amino acids, 58-60 amino acids, or 60-65 amino acids.

[0066] In some of the optional embodiments, the encoded peptide linker comprises polypeptides having a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65 amino acids. In some of the optional embodiments, the encoded peptide linker comprises GS, GGS, GGGGS (SEQ ID NO: 43), GGGGGS (SEQ ID NO: 41), and combinations thereof. In some of the optional embodiments, the encoded peptide linker includes (GGS)n (where n is 1 to 10). In some of the optional embodiments, the encoded peptide linker includes (GGGGS)n (SEQ ID NO: 42) (where n is 1 to 10). In some of the optional embodiments, the encoded peptide linker includes (GGGGGS)n (SEQ ID NO: 27) (where n is 1 to 4). In some of the optional embodiments, the sequence encoding the G protein is wild-type Nipah virus G (NiV-G) protein or Hendra virus G protein, or a functionally active variant or biologically active moiety thereof. In some embodiments, the variant is a variant of that which exhibits reduced binding for its native binding partner. In some of the optional embodiments, the nucleic acid sequence encoding the G protein is wild-type Nipah virus G (NiV-G) protein or Hendra virus G protein, or a variant of that which exhibits reduced binding for its native binding partner. In some embodiments, the encoded G protein is the wild-type NiV-G protein or a functionally active variant or biologically active portion thereof. In some of the embodiments, the nucleic acid sequence encoding the G protein is the wild-type NiV-G protein. In some of the embodiments, the nucleic acid sequence encoding the G protein is a mutant NiV-G protein exhibiting reduced binding to ephrin B2 or ephrin B3.

[0067] In some of the embodiments, the NiV-G protein or its functionally active variant or biologically active moiety comprises the amino acid sequence shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, or comprises at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or less The amino acid sequence contains at least 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99% sequence identity. In some of the various embodiments, the NiV-G protein is a cleaved and biologically active portion lacking up to 40 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44).In some of the arbitrary embodiments, the NiV-G protein is a biologically active portion cleaved at the N-terminus of wild-type NiV-G, which is the sequence shown in any of SEQ ID NOs. 10-15, 35-40 or 45-50, or at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, and It contains an amino acid sequence having sequence identity of at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%.

[0068] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that includes a 5-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 10. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 35.In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 45.

[0069] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that includes a 10-amino acid cleavage at or near its N-terminus. In some of the embodiments, the mutant NiV-G protein or biologically active moiety includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 11. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 36.In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 46.

[0070] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that includes a 15-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 12. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 37.In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 47.

[0071] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that includes a 20-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 13. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 38.In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 48.

[0072] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that includes a 25-amino acid cleavage at or near the N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 14. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 39.In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 49.

[0073] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that includes a 30-amino acid cleavage at or near the N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 15. In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 40.In some of the embodiments, the NiV-G protein or the biologically active portion includes an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 50.

[0074] In some of the optional embodiments, the NiV-G protein is a biologically active portion of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) that has a 34-amino acid cleavage at or near its N-terminus. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 22. In some of the embodiments, the NiV-G protein or the biologically active portion has an amino acid sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 53.

[0075] In some of the embodiments, the G protein is a mutant NiV-G protein exhibiting reduced binding to ephrin B2 or ephrin B3. In some of the embodiments, the mutant NiV-G protein contains one or more amino acid substitutions corresponding to amino acid substitutions selected from the group consisting of E501A, W504A, Q530A and E533A, with reference to the numbering shown in Sequence ID No. 28. In some of the embodiments, the mutant NiV-G protein contains amino acid substitutions E501A, W504A, Q530A and E533A, with reference to the numbering shown in Sequence ID No. 28.

[0076] In some of the various embodiments, the mutant NiV-G protein includes i) a cleavage at or near the N-terminus, and ii) a point mutation selected from the group consisting of E501A, W504A, Q530A, and E533A. In some of the various embodiments, the mutant NiV-G protein contains an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 16. In some of the embodiments, the mutant NiV-G protein contains an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 51.

[0077] In some of the optional embodiments, the F protein or its biologically active portion is the wild-type Nipah virus F (NiV-F) protein or the Hendra virus F protein, or a functionally active variant or biologically active portion thereof. In some of the embodiments, the NiV-F protein or its functionally active variant or biologically active moiety includes the amino acid sequence shown in SEQ ID NO: 2, or an amino acid sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 2.

[0078] In some of the optional embodiments, the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 20-amino acid cleavage at or near its C-terminus. In some of the embodiments, the NiV-F protein or the biologically active moiety has an amino acid sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 5. In some of the optional embodiments, the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) that includes i) a 20-amino acid cleavage at or near the C-terminus, and ii) a point mutation at the N-linked glycosylation site.

[0079] In some of the embodiments, the NiV-F protein or the biologically active portion has an amino acid sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 7.

[0080] In some of the optional embodiments, the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 22-amino acid cleavage at or near its C-terminus. In some of the various embodiments, the NiV-F protein or the biologically active portion has an amino acid sequence encoded by a sequence of nucleotides encoding the sequence shown in SEQ ID NO: 8, or a sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 8.

[0081] In some of the various embodiments, the NiV-F protein has an amino acid sequence having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 23. In some of the embodiments, the F protein comprises the sequence shown in SEQ ID NO: 23, and the G protein comprises the sequence shown in SEQ ID NO: 16.

[0082] This specification provides vectors comprising polynucleotides of any of the embodiments described herein. In some of the embodiments, the vector is a mammalian vector, a viral vector, or an artificial chromosome, and optionally the artificial chromosome is a bacterial artificial chromosome (BAC).

[0083] This specification provides plasmids comprising polynucleotides of any of the embodiments described herein. In some of the embodiments, the plasmid further comprises one or more nucleic acids encoding proteins for lentiviral generation.

[0084] This specification provides cells comprising a polynucleotide of any embodiment described herein, a vector of any embodiment described herein, or a plasmid of any embodiment described herein.

[0085] This specification provides a method for producing targeted lipid particles comprising a henipavirus F protein molecule or its biologically active portion, and a targeted envelope protein comprising a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and a single-domain antibody (sdAb) variable domain, a) To provide a cell comprising a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and a single-domain antibody (sdAb) variable domain. b) Culturing cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying targeted lipid particles from cells, thereby producing targeted lipid particles. A method including this is provided.

[0086] This specification describes a method for producing a pseudotyped lentiviral vector, a) To provide producer cells comprising lentiviral nucleic acids (multiple), a nucleic acid encoding a henipavirus F protein molecule or its biologically active portion, and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and a single-domain antibody. b) Culturing cells under conditions that enable the generation of lentiviral vectors, and c) Isolating, concentrating, or purifying lentiviral vectors from cells, thereby producing pseudotyped lentiviral vectors. A method including this is provided.

[0087] In some of the optional embodiments, the single-domain antibody binds to a cell surface molecule present on the target cell. In some of the optional embodiments, the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule. In some of the optional embodiments, the target cell is selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. In some of the optional embodiments, the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytic cells, SLC7A10+ adipocytes, or CD30+ lung epithelial cells. In some of the optional embodiments, the single-domain antibody binds to an antigen or a portion thereof present on the target cells.

[0088] This specification describes a method for producing targeted lipid particles comprising a henipavirus F protein molecule or its biologically active portion, and a targeted envelope protein comprising a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and binding domain, a) To provide a cell comprising a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion and a binding domain, and the binding domain binds to (i) a cell surface molecule selected from the group consisting of ASGR1, ASGR2, and TM4SF5, optionally human ASGR1, human ASGR2, and human ASGR2; (ii) a cell surface molecule selected from the group consisting of CD4 or CD8, optionally human CD4 or human CD8; or (iii) a cell surface molecule that is a low-density lipoprotein receptor (LDL-R), optionally human LDL-R. b) Culturing cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying targeted lipid particles from cells, thereby producing targeted lipid particles. A method including this is provided.

[0089] This specification provides a method for producing a pseudotyped lentiviral vector, comprising: a) lentiviral nucleic acid(s); a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof; and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion and a binding domain, and the binding domain is (i) ASGR1, ASGR2, and TM4SF5, optionally human ASGR1, human ASG A method is provided comprising: a) providing a cell surface molecule that binds to (ii) a cell surface molecule selected from the group consisting of R2 and human ASGR2; b) a cell surface molecule that binds to CD4 or CD8, optionally selected from the group consisting of human CD4 or human CD8; or c) a cell surface molecule that binds to a low-density lipoprotein receptor (LDL-R), optionally human LDL-R; b) culturing producer cells under conditions that enable the production of a lentiviral vector; and c) isolating, concentrating, or purifying a lentiviral vector from cells to produce a pseudotyped lentiviral vector.

[0090] In some of the embodiments, the binding domain is a single-domain antibody. In some of the embodiments, the binding domain is a single-stranded variable fragment (scFv). In some of the embodiments, the cell surface molecule is selected from the group consisting of ASGR1, ASGR2, and TM4SF5. In some of the embodiments, the cell surface molecule is CD8 or CD4. In some of the embodiments, the cell surface molecule is LDL-R.

[0091] This specification provides a method for producing targeted lipid particles comprising a henipavirus F protein molecule or a biologically active portion thereof and a targeted envelope protein, a) To provide cells comprising a polynucleotide of any embodiment provided herein, a vector of any embodiment described herein, or a plasmid of any embodiment described herein. b) Culturing cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying targeted lipid particles from cells, thereby producing targeted lipid particles. A method including this is provided.

[0092] This specification describes a method for producing a pseudotyped lentiviral vector, a) To provide producer cells comprising lentiviral nucleic acid(s) and a polynucleotide of any of the embodiments listed herein or a vector of any of the embodiments listed herein, b) Culturing cells under conditions that enable the generation of lentiviral vectors, and c) Isolating, concentrating, or purifying lentiviral vectors from cells, thereby producing pseudotyped lentiviral vectors. A method is provided which includes the following. In some of the optional embodiments, prior to step (b), the method further includes providing cells with a henipavirus F protein molecule or a polynucleotide encoding a biologically active portion thereof.

[0093] In some of the arbitrary embodiments, the cells are mammalian cells.

[0094] In some of the optional embodiments, the cells are producer cells containing viral nucleic acids. In some of the optional embodiments, the viral nucleic acids are retroviral nucleic acids or lentiviral nucleic acids, and the targeted lipid particles are viral particles or virus-like particles. In some of the optional embodiments, the viral particles or virus-like particles are retroviral particles or retroviral-like particles. In some embodiments, the viral particles or virus-like particles are lentiviral particles or lentiviral-like particles.

[0095] In some of the optional embodiments, the viral nucleic acid(s) lacks one or more genes involved in viral replication. In some of the optional embodiments, the viral nucleic acid comprises nucleic acid encoding a viral packaging protein selected from one or more of Gag, Pol, Rev, and Tat. In some of the optional embodiments, the viral nucleic acid comprises one or more (e.g., all) of the following nucleic acid sequences: 5'LTR (e.g., containing U5 and lacking a functional U3 domain), pseudopackaging element (psi), central polypurine sequence (cPPT) / central termination sequence (CTS) (e.g., DNA flap), poly-A tail sequence, post-transcriptional regulatory element (e.g., WPRE), Rev reaction element (RRE), and 3'LTR (e.g., containing U5 and lacking a functional U3).

[0096] This specification provides producer cells comprising a polynucleotide of any of the embodiments listed herein, a vector of any of the embodiments listed herein, or a plasmid of any of the embodiments described herein.

[0097] In some of the optional embodiments, the producer cells further include nucleic acids that encode the henipavirus F protein or a biologically active portion thereof.

[0098] In some of the optional embodiments, the cell further comprises viral nucleic acid. In some of the optional embodiments, the viral nucleic acid is lentiviral nucleic acid. This specification provides a producer cell comprising (i) viral nucleic acid(s), (ii) a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and (iii) a nucleic acid encoding a targeted envelope protein comprising henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and a single-domain antibody (sdAb) variable domain, wherein the viral nucleic acid(s) is optionally lentiviral nucleic acid. In some of the optional embodiments, the single-domain antibody binds to a cell surface molecule present on the target cell. In some of the optional embodiments, the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule.

[0099] In some of the optional embodiments, the target cells are selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. In some of the optional embodiments, the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytic cells, SLC7A10+ adipocytes, or CD30+ lung epithelial cells. In some of the optional embodiments, the single-domain antibody binds to an antigen or a portion thereof present on the target cells.

[0100] This specification provides producer cells comprising (i) viral nucleic acids(or more), (ii) nucleic acids encoding a henipavirus F protein molecule or a biologically active portion thereof, and (iii) nucleic acids encoding a targeted envelope protein including a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion and a binding domain thereof, wherein the binding domain binds to (i) cell surface molecules selected from the group consisting of ASGR1, ASGR2, and TM4SF5, optionally human ASGR1, human ASGR2, and human ASGR2, (ii) cell surface molecules selected from the group consisting of CD4 or CD8, optionally human CD4 or human CD8, or (iii) cell surface molecules that are low-density lipoprotein receptors (LDL-R), optionally human LDL-R. In some of the various embodiments, the viral nucleic acids(or more) are lentiviral nucleic acids.

[0101] In some of the optional embodiments, the cell surface molecule or antigen is selected from the group consisting of ASGR1, ASGR2, and TM4SF5. In some of the optional embodiments, the cell surface molecule or antigen is CD8 or CD4. In some of the optional embodiments, the cell surface molecule or antigen is LDL-R.

[0102] In some of the optional embodiments, the viral nucleic acid(s) lacks one or more genes involved in viral replication. In some of the optional embodiments, the viral nucleic acid comprises a nucleic acid encoding a viral packaging protein selected from one or more of Gag, Pol, Rev, and Tat.

[0103] In some of the various embodiments, the viral nucleic acid comprises one or more (e.g., all) of the following nucleic acid sequences: 5'LTR (e.g., containing U5 and lacking a functional U3 domain), psipackaging elements (psi), central polypurine sequences (cPPT) / central termination sequences (CTS) (e.g., DNA flap), poly(A) tail sequences, post-transcriptional regulatory elements (e.g., WPRE), Rev reaction elements (RRE), and 3'LTR (e.g., containing U5 and lacking a functional U3).

[0104] In some of the embodiments, the henipavirus F protein molecule or its biologically active portion comprises (i) the sequence shown in SEQ ID NO: 2, and (ii) an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 2. In some of the various embodiments, the henipavirus F protein molecule or its biologically active portion comprises (i) the sequence shown in SEQ ID NO: 5, and (ii) an amino acid sequence having at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 5.

[0105] In some of the embodiments, the henipavirus F protein molecule or its biologically active portion comprises (i) the sequence shown in SEQ ID NO: 7, and (ii) an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 7. In some of the embodiments, the henipavirus F protein molecule or its biologically active portion is (i) a sequence encoded by a nucleotide sequence encoding the sequence shown in SEQ ID NO: 8, (ii) at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or It comprises an amino acid sequence encoded by a nucleotide sequence encoding a sequence having approximately 88%, or at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0106] In some of the embodiments, the henipavirus F protein molecule or its biologically active portion is (i) the sequence shown in SEQ ID NO: 23, and (ii) SEQ ID NO: 23 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88%. It contains amino acid sequences having sequence identity of approximately 88%, or at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0107] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, or (ii) SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44 in proportion to at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or less It also contains amino acid sequences having sequence identity of 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0108] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 10, (ii) SEQ ID NO: 10 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0109] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 35, (ii) SEQ ID NO: 35 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0110] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 45, (ii) SEQ ID NO: 45 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0111] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 11, (ii) SEQ ID NO: 11 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0112] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 36, (ii) SEQ ID NO: 36 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0113] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 46, (ii) SEQ ID NO: 46 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0114] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 12, (ii) SEQ ID NO: 12 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0115] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 37, (ii) SEQ ID NO: 37 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0116] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 47, (ii) SEQ ID NO: 47 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0117] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 13, (ii) SEQ ID NO: 13 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0118] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 38, (ii) SEQ ID NO: 38 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0119] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 48, (ii) SEQ ID NO: 48 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0120] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 14, (ii) SEQ ID NO: 14 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0121] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 39, (ii) SEQ ID NO: 39 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0122] In some of the embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 49, (ii) SEQ ID NO: 49 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0123] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 15, (ii) SEQ ID NO: 15 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0124] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 40, (ii) SEQ ID NO: 40 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0125] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 50, (ii) SEQ ID NO: 50 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0126] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 16, (ii) SEQ ID NO: 16 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0127] In some of the various embodiments, the henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion is (i) the sequence shown in SEQ ID NO: 51, (ii) SEQ ID NO: 51 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. It contains amino acid sequences having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, at least 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0128] In some aspects of the provided embodiments, the targeted lipid particles have more targeted envelope protein expression compared to reference lipid particles in which the same envelope protein is incorporated into a similar lipid bilayer but is fused to an alternative targeting site, and optionally the alternative targeting site is a single-chain variable fragment (scFv). In some of the embodiments, the expression increases by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 400%, 500% or more or above that. In some embodiments, the expression increases by 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold or more or above that, preferably by 10-fold or about 10-fold or about 10-fold more or above that. In some of the embodiments, the titer in the target cells after transduction is 1×10 6 transducing units (TU) / mL or more, 2×10 6 TU / mL or more, 3×10 6 TU / mL or more, 4×10 6 TU / mL or more, 5×10 6 TU / mL or more, 6×10 6 TU / mL or more, 7×10 6 TU / mL or more, 8×10 6 TU / mL or more, 9×10 6 TU / mL or more, or 1×10 7 TU / mL or more. Also, herein, compositions are provided in which 50% or about 50% or more, 55% or about 55% or more, 60% or about 60% or more, 65% or about 65% or more, 70% or about 70% or more, or 75% or about 75% or more of the population of lipid particles are surface positive for the targeted envelope protein. In some of the embodiments, the targeted envelope protein is present on the surface of the targeted lipid particles at a density of at least about (0.001, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2 or 0.5) of the targeted envelope protein / nm 2 of the targeted lipid particles.

[0129] This specification provides viral vector particles or virus-like particles generated from producer cells of any of the embodiments provided herein.

[0130] This specification provides compositions comprising a plurality of targeted lipid particles of any of the embodiments provided herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some of the embodiments, the targeted lipid particles have an average diameter of less than 1 μm. In some of the embodiments, the composition contains at least about (0.001, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, or 0.5) targeted envelope proteins / nm 2 It further contains targeted envelope proteins present on the surface of targeted lipid particles at an average density.

[0131] This specification provides producer cells containing membrane (e.g., cell membrane) expression of a targeted envelope protein, which is more abundant than in reference producer cells in which the same envelope protein is incorporated into the membrane (e.g., cell membrane) but is fused to an alternative targeting site, wherein the alternative targeting site is optionally a single-stranded variable fragment (scFv). In some embodiments, the expression is increased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 400%, 500% or more. In some embodiments, the expression is increased by 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times, 30 times or more, preferably 10 times or about 10 times or more than 10 times. In some embodiments, the producer cells have expression of a targeted envelope protein on the producer cell membrane (e.g., cell membrane) of at least 20 proteins per square micron (e.g., at least 50, 100, 200, 500, 1000, 2000, 5000, or 10,000 proteins). In some of the arbitrary embodiments, the targeted envelope protein constitutes at least 0.1% (e.g., at least 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%) of the total membrane (e.g., cell membrane) proteins of the producer cells (e.g., relative to total protein weight).

[0132] This specification provides a method for transducing cells, comprising transducing cells with any of the viral vectors or compositions described herein. In some of the optional embodiments, the targeted envelope protein of the lentiviral vector or targeted lipid particle targets CD4, and the cells are CD4+ cells. In some of the optional embodiments, the targeted envelope protein of the lentiviral vector targets CD8, and the cells are CD8+ cells. In some of the optional embodiments, the targeted envelope protein of the lentiviral vector targets ASGR1, ASGR2, or TM4SF5, and the cells are hepatocytes.

[0133] This specification provides a method for delivering an exogenous substance to a subject (e.g., a human subject), comprising administering targeted lipid particles or a composition of any of the embodiments provided herein to the subject, wherein the targeted lipid particles or lentiviral vector contain the exogenous substance.

[0134] This specification provides a method for delivering an exogenous substance to a subject (e.g., a human subject), comprising administering one of the compositions described herein to the subject, wherein a plurality of targeted lipid particles or lentiviral vectors contain the exogenous substance.

[0135] This specification provides a method for delivering a chimeric antigen receptor (CAR) to a cell, comprising contacting the cell with a targeted lipid particle of any of the lentiviral vectors described herein or any of the embodiments described herein, wherein the lentiviral vector or targeted lipid particle comprises a nucleic acid encoding the CAR.

[0136] This specification provides a method for delivering a chimeric antigen receptor (CAR) to a cell, comprising contacting the cell with any of the compositions described herein, wherein a plurality of lentiviral vectors or targeted lipid particles contain nucleic acids encoding the CAR.

[0137] This specification provides a method for delivering an exogenous substance to hepatocytes, comprising contacting the cells with any of the lentiviral vectors described herein or any of the embodiments described herein, or with targeted lipid particles or lentiviral vectors.

[0138] This specification provides a method for delivering an exogenous substance to hepatocytes, comprising contacting the cells with one of the compositions described herein, wherein a plurality of lentiviral vectors or targeted lipid particles contain the exogenous substance for delivery to hepatocytes. In some of the optional embodiments, the contact is transduction of the cells with the lentiviral vectors or targeted lipid particles.

[0139] This specification provides a method for treating a disease or disorder in a subject (e.g., a human subject), comprising administering targeted lipid particles of any of the embodiments provided herein or a composition of any of the embodiments provided herein to the subject.

[0140] This specification provides a method for fusing mammalian cells to targeted lipid particles, comprising administering the targeted lipid particles of any embodiment provided herein or a composition of any embodiment provided herein to a subject. In some of the optional embodiments, fusing mammalian cells to targeted lipid particles delivers an exogenous substance to a subject (e.g., a human subject). In some of the optional embodiments, fusing mammalian cells to targeted lipid particles treats a disease or disorder in a subject (e.g., a human subject). In some of the optional embodiments, the targeted envelope protein of the lentiviral vector or targeted lipid particle targets CD4, and the cell is a CD4+ cell. In some of the optional embodiments, the targeted envelope protein of the lentiviral vector targets CD8, and the cell is a CD8+ cell. In some of the optional embodiments, the targeted envelope protein of the lentiviral vector targets ASGR1, ASGR2, or TM4SF5, and the cell is a hepatocyte.

[0141] In some of the optional embodiments, the targeted lipid particles have higher targeted envelope protein expression compared to reference lipid particles in which the same envelope protein is incorporated into a similar lipid bilayer but is fused to an alternative targeting site. In some embodiments, the alternative targeting site is a single-stranded variable fragment (scFv). In some of the optional embodiments, the expression increases by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 400%, 500% or more. In some of the optional embodiments, the expression increases by 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times, 30 times or more, preferably 10 times or about 10 times or more than 10 times.

[0142] In some of the optional embodiments, the titer in the target cells after transduction is 1 × 10⁻⁶. 6 Transduction units (TU) / mL or more, 2 × 10 6TU / mL or more, 3×10 6 TU / mL or more, 4×10 6 TU / mL or more, 5×10 6 TU / mL or more, 6×10 6 TU / mL or more, 7×10 6 TU / mL or more, 8×10 6 TU / mL or more, 9×10 6 TU / mL or higher, or 1 × 10 7 It is TU / mL or higher.

[0143] In some of the optional embodiments, within the population of lipid particles or lentiviral vectors in the composition, 50% or more than 50%, 55% or more than 55%, 60% or more than 60%, 65% or more than 65%, 70% or more than 70%, or 75% or more than 75% are surface-positive for the targeted envelope protein. In some of the optional embodiments, the targeted envelope protein is at least about (0.001, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, or 0.5) of targeted envelope protein / nm 2 It is present on the surface of the targeted lipid particles at a density of [value].

[0144] This specification describes a composition comprising a plurality of targeted lipid particles of any embodiment described herein or a plurality of lentiviral vectors of any embodiment described herein, wherein the targeted envelope protein is at least about (0.001, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, or 0.5) of targeted envelope protein / nm. 2 A composition is provided that exists on the surface of targeted lipid particles at an average density.

[0145] In some of the arbitrary embodiments, the producer cells have membrane (e.g., cell membrane) expression of the targeted envelope protein, which is more abundant than in the reference producer cells, where the same envelope protein is incorporated into the membrane (e.g., cell membrane), but it is fused to an alternative targeting site, and the alternative targeting site is optionally a single-stranded variable fragment (scFv). In some of the arbitrary embodiments, the expression increases by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 400%, 500% or more. In some of the optional embodiments, the expression increases by 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times, 30 times or more, preferably 10 times or about 10 times or more. In some of the optional embodiments, the producer cells have expression of a targeted envelope protein on the membrane (e.g., cell membrane) of the producer cells, which is at least 20 proteins per square micron (e.g., at least 50, 100, 200, 500, 1000, 2000, 5000, or 10,000 proteins). In some of the arbitrary embodiments, the targeted envelope protein constitutes at least 0.1% (e.g., at least 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%) of the total membrane (e.g., cell membrane) proteins of the producer cell (e.g., relative to the total protein weight).

[0146] Detailed explanation This specification provides targeted lipid particles comprising a lipid bilayer surrounding a lumen or cavity, and (1) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof, and (2) a targeted envelope protein containing a binding domain, e.g., a single-domain antibody (sdAb) variable domain, wherein the targeted envelope protein is embedded in the lipid bilayer of the lipid particle. In certain embodiments, the binding domain, e.g., the single-domain antibody, is an antibody that binds to a desired target molecule, e.g., an antibody having the ability to bind specifically. Exemplary binding domains are described in Section II.A.2. In some embodiments, the targeted lipid particles also contain a henipavirus fusion (F) protein molecule or a biologically active moiety thereof embedded in the lipid bilayer. In certain embodiments, the lipid particles may be virus-like particles, viruses, or viral vectors, e.g., lentiviral vectors.

[0147] In some embodiments, one or both of the G protein and / or F protein are derived from the Hendra (HeV) or Nipah (NiV) virus, or are their biologically active portions, variants, or variants. In certain embodiments, both the G protein and / or F protein are derived from the Hendra (HeV) or Nipah (NiV) virus. In some embodiments, the fusion and binding glycoproteins mediate the entry of Nipah virus into cells.

[0148] F proteins, such as NiV-F, are class I fusion proteins that share structural and functional features with many families of fusion proteins (e.g., HIV-1 gp41 or influenza virus hemagglutinin [HA]), including hydrophobic fusion peptides and ectodomains with two heptad repeat regions (White JM et al. 2008. Crit Rev Biochem Mol Biol 43:189-219). F proteins are synthesized as an inactive precursor F0 and activated by proteolytic cleavage into two disulfide-linked subunits F1 and F2 (Moll M. et al. 2004. J. Virol. 78(18):9705-9712).

[0149] The G protein is a type II transmembrane glycoprotein containing an N-terminal cytoplasmic tail, transmembrane domain, extracellular stalk, and bulbous head, and is the binding protein for henipavirus (e.g., Nipah virus or Hendra virus) (Liu, Q. et al. 2015. Journal of Virology, 89(3):1838-1850). The binding protein NiV-G recognizes receptors ephrin B2 and ephrin B3. Receptor binding to NiV-G triggers a series of structural changes that ultimately lead to NiV-F activation, exposing the NiV-F fusion peptide and enabling another series of structural changes that result in viral cell membrane fusion (Stone JA et al. 2016. J Virol. 90(23):10762-10773). Ephrin B2 was previously identified as the primary NiV receptor (Negrete et al., 2005), and ephrin B3 as an alternative receptor (Negrete et al., 2006). In fact, NiV-G exhibits high affinity for ephrin B2 and B3 with picomolar affinity binding constants (Kd) (Negrete et al., 2006) (Kd = 0.06 nM and 0.58 nM, respectively, for ephrin B2 and B3 expressed on the cell surface).

[0150] The efficiency of transduction of targeted lipid particles can be improved by manipulating superfusion mutations in one or both NiV-F and NiV-G. Several such mutations have been previously described (see, e.g., Lee et al, 2011, Trends in Microbiology). This may be useful, for example, to maintain the specificity and picomolar affinity of NiV-G to ephrin B2 and / or B3. Mutations in NiV-G that completely inactivate ephrin B2 and B3 binding but do not affect the association of NiV-G with NiV-F have also been identified. Methods to improve the targeting of lipid particles can be achieved by fusing a binding molecule with a G protein (e.g., Niv-G containing a Niv-G with a mutation to inactivate ephrin B2 and ephrin B3 binding). This may enable modified G protein directivity that allows targeting of other desired cell types other than ephrin B2+ through the addition of binding molecules directed towards different cell surface molecules.

[0151] Retargeted lipid particles incorporating such binding molecules fused to G proteins have been generated, and it has been found herein that some binding molecules are expressed better on the surface of the lipid particles than others when fused to G proteins (e.g., NiV-G). For example, single-domain antibodies (sdAbs), e.g., VHH, have been found to be expressed 10 times better than single-stranded variable fragments (scFv). While we do not wish to be bound by theory, the increased expression may be attributable to the increased stability of the retargeted G protein on the surface of the lipid particle. This greater expression may improve the ability of the lipid particles to target molecules (e.g., cell surface molecules) compared to similar lipid particles containing alternative binding domains for the same target molecule, e.g., scFv.

[0152] Accordingly, this specification provides targeted lipid particles containing a henipavirus G protein (e.g., Hendra or Nipah, e.g., NiV-G) conjugated to an sdAb variable domain that is directed to or capable of binding to cell surface molecules on target cells. The sdAb variable domain may include sdAb of VL or VH only, nanobodies, camelid VHH domains, shark IgNAR or fragments thereof. In some embodiments, the sdAb is VHH.

[0153] In aspects of the provided embodiments, targeted lipid particles may be engineered to express a henipavirus F protein molecule or its biologically active moiety, and a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and (ii) a single-domain antibody (sdAb) variable domain, wherein the F protein molecule or its biologically active moiety and the targeted envelope protein are embedded in a lipid bilayer. In some embodiments, the sdAb variable domain is conjugated to the C-terminus of the G protein or its biologically active moiety. In some embodiments, the sdAb variable domain is conjugated to the G protein via a linker.

[0154] Furthermore, targeted lipid particles are provided that additionally contain one or more exogenous substances, including after in vivo administration to a target, for example, for the delivery of diagnostic or therapeutic agents to cells. Also provided herein are methods and uses of targeted lipid particles in diagnostic and therapeutic methods. Also provided are methods for manipulating, preparing, and generating polynucleotides, targeted lipid noncellular particles, compositions containing particles, and kits and devices for using, generating, and administering the particles.

[0155] All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated by reference in whole for all purposes to the same extent that each individual publication is disclosed by reference. If any definition provided herein contradicts or otherwise conflicts with any definition provided herein in a patent, application, published application, or other publication incorporated herein by reference, the definition provided herein shall prevail over the definition incorporated herein by reference.

[0156] The section headings used herein are for organizational purposes only and should not be construed as limiting the subjects described. [Invention 1001] Targeted lipid particles, (a) A lipid bilayer surrounding the lumen, (b) Henipavirus F protein molecule or its biologically active portion, (c) A targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and (ii) a single-domain antibody (sdAb) variable domain, wherein the sdAb variable domain is bound to the C-terminus of the G protein or its biologically active moiety, and / or the sdAb is bound to the G protein or its biologically active moiety via a peptide linker, and the sdAb binds to a cell surface molecule of a target cell. The targeted lipid particle comprises the F protein molecule or its biologically active portion and the targeted envelope protein, wherein the F protein molecule or its biologically active portion and the targeted envelope protein are embedded in a lipid bilayer. [Invention 1002] Targeted lipid particles of the present invention 1001, wherein the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule. [Invention 1003] Targeted lipid particles of the present invention 1001 or 1002, wherein the target cells are selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. [Invention 1004] Targeted lipid particles according to any of invention 1001 to 1003, wherein the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytocytes, SLC7A10+ adipocytes, or CD30+ lung epithelial cells. [Invention 1005] A targeted lipid particle according to any of the present invention 1001 to 1004, wherein the single-domain antibody binds to an antigen or a portion thereof present on hepatocytes. [Invention 1006] Targeted lipid particles according to any of the present invention 1001 to 1005, wherein the cell surface molecule or antigen is selected from the group consisting of ASGR1, ASGR2, and TM4SF. [Invention 1007] A targeted lipid particle according to any of the present invention 1001 to 1004, wherein the single-domain antibody binds to an antigen or a part thereof present on a T cell. [Invention 1008] Targeted lipid particles according to any one of the present invention 1001 to 1004 and 1007, wherein the cell surface molecule or antigen is CD8 or CD4. [Invention 1009] A targeted lipid particle according to any one of the present invention 1001 to 1004, wherein the cell surface molecule or antigen is LDL-R. [Invention 1010] Targeted lipid particles, (a) A lipid bilayer surrounding the lumen, (b) Henipavirus F protein molecule or its biologically active portion, (c) A targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active moiety, and the binding domain binds to a cell surface molecule selected from the group consisting of ASGR1, ASGR2, TM4SF5, CD8, CD4 and LDL-R. A targeted lipid particle comprising the F protein molecule or its biologically active portion and the targeted envelope protein, wherein the F protein molecule or its biologically active portion and the targeted envelope protein are embedded in the lipid bilayer. [Invention 1011] The targeted lipid particle of the present invention 1010, wherein the binding domain is bound to the G protein via a linker. [Invention 1012] The targeted lipid particle of the present invention 1011, wherein the linker is a peptide linker. [Invention 1013] A lentiviral vector, which is a targeted lipid particle according to any of invention 1001 to 1012. [Invention 1014] (a) Henipavirus F protein molecule or its biologically active portion, (b) A targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active moiety, and the binding domain binds to CD4, (c) A cargo comprising a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR is (i) an extracellular antigen-binding domain that binds to CD19, wherein the extracellular antigen-binding domain is optionally scFv, (ii) transmembrane domain, and (iii) Intracellular signaling domain including the CD3 zeta signaling domain Including the cargo and A lentiviral vector containing... [Invention 1015] The lentiviral vector of the present invention 1014, wherein the intracellular signaling region of the CAR further comprises a 4-1BB costimulatory signaling domain. [Invention 1016] The lentiviral vector of the present invention 1014 or 1015 is capable of delivering the nucleic acid encoding the CAR to T cells, and optionally the T cells are in vivo in the subject. [Invention 1017] (a) Henipavirus F protein molecule or its biologically active portion, (b) A targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or its biologically active moiety, and the binding domain binds to a cell surface molecule selected from the group consisting of ASGR1, ASGR2 and TM4SF5. A lentiviral vector containing... [Invention 1018] A lentiviral vector according to the present invention 1017, capable of targeting liver cells. [Invention 1019] A lentiviral vector according to either Invention 1017 or Invention 1018, further comprising an exogenous substance for delivery to hepatocytes, and / or the lentiviral vector is capable of delivering the exogenous substance to hepatocytes, wherein the hepatocytes are optionally in vivo in the subject. [Invention 1020] A lentiviral vector according to any of the invention 1014 to 1019, wherein the binding domain is bound to the G protein via a linker, and the linker is optionally a peptide linker. [Invention 1021] A targeted lipid particle according to any of Invention 1010 to 1013 or a lentiviral vector according to any of Invention 1014 to 1020, wherein the binding domain is a single-domain antibody or a single-stranded variable fragment (scFv). [Invention 1022] The peptide linker comprises a peptide linker having a length of up to 65 amino acids, and optionally a length of 2 to 65 amino acids, and is a targeted lipid particle according to any of Invention 1001-1009, 1012, and 1013, or a lentiviral vector according to Invention 1020 or Invention 1021. [Invention 1023] The peptide linker is a flexible linker containing GS, GGS, GGGGS (SEQ ID NO: 43), GGGGGS (SEQ ID NO: 41), or a combination thereof, or the peptide linker contains (GGS)n where n is 1 to 10, (GGGGS)n where n is 1 to 10 (SEQ ID NO: 42), or (GGGGGS)n where n is 1 to 6 (SEQ ID NO: 27), wherein the targeted lipid particles are any of Invention 1001 to 1009, 1012, 1013, and 1022, or a lentiviral vector is any of Invention 1020 to 1022. [Invention 1024] A targeted lipid particle according to any of Invention 1001-1013, 1022, and 1023, or a lentiviral vector according to any of Invention 1014-1023, wherein the G protein or its biologically active portion is wild-type Nipah virus G (NiV-G) protein or Hendra virus G protein, or a functionally active variant or biologically active portion thereof. [Invention 1025] A targeted lipid particle according to any of Invention 1001-1013 and 1022-1024, or a lentiviral vector according to any of Invention 1014-1024, wherein the G protein or its biologically active portion is a wild-type NiV-G protein or a functionally active variant or biologically active portion thereof. [Invention 1026] The NiV-G protein or its functionally active variant or biologically active moiety is at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 88%. Targeted lipid particles or lentiviral vectors of the present invention 1024 or 1025, comprising an amino acid sequence having sequence identity of approximately 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1027] The aforementioned NiV-G protein, A cleaved and biologically active moiety lacking up to 40 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44). A targeted lipid particle or lentiviral particle according to any of the present invention 1024 to 1026. [Invention 1028] The NiV-G protein is a biologically active portion cleaved at the N-terminus of wild-type NiV-G, and is a sequence shown in any of SEQ ID NOs. 10-15, 35-40, or 45-50, or at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 8 Targeted lipid particles or lentiviral particles according to any of the present invention 1024 to 1027, having an amino acid sequence having sequence identity of 8% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1029] The NiV-G protein is a biologically active moiety having a cleavage at or near the N-terminus of the wild-type NiV-G protein, selected from the group consisting of a 5-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 10-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 15-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 20-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 25-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 30-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, or a 34-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, wherein the wild-type NiV-G protein is optionally a targeted lipid particle or lentiviral vector of any of the present invention 1024-1028 as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44. [Invention 1030] A targeted lipid particle or lentiviral vector according to any of the present invention 1024-1029, wherein the NiV-G protein is a biologically active portion having a 34-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44). [Invention 1031] A targeted lipid particle or lentiviral vector according to any of Invention 1001-1013 and 1022-1030, wherein the G protein or its biologically active portion is a mutant NiV-G protein exhibiting reduced binding to ephrin B2 or ephrin B3. [Invention 1032] The aforementioned mutant NiV-G protein, Refer to the numbering shown in Sequence ID No. 28 and select one or more amino acid substitutions corresponding to amino acid substitutions selected from the group consisting of E501A, W504A, Q530A, and E533A. A targeted lipid particle or lentiviral vector according to the present invention 1031, comprising: [Invention 1033] The aforementioned mutant NiV-G protein, Refer to the numbering shown in Sequence ID No. 28 for amino acid substitutions E501A, W504A, Q530A, and E533A. A targeted lipid particle or lentiviral vector according to Invention 1031 or Invention 1032, comprising: [Invention 1034] The mutant NiV-G protein or the biologically active portion is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, less A targeted lipid particle or lentiviral vector according to any of the present invention 1031 to 1033, having an amino acid sequence having sequence identity of at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1035] A targeted lipid particle or lentiviral vector according to any of Invention 1001-1013 and 1022-1034, wherein the F protein or its biologically active portion is wild-type Nipah virus F (NiV-F) protein or Hendra virus F protein, or a functionally active variant or biologically active portion thereof. [Invention 1036] A targeted lipid particle according to any of invention 1001-1013 and 1022-1035, or a lentiviral vector according to any of invention 1014-1035, wherein the F protein or its biologically active portion is a wild-type NiV-F protein or a functionally active variant or biologically active portion thereof. [Invention 1037] The NiV-F protein or its functionally active variant or biologically active moiety is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, or at least 90%. Targeted lipid particles of any of Invention 1001-1013 and 1022-1036 or lentiviral vectors of any of Invention 1014-1036, comprising an amino acid sequence having sequence identity of approximately 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1038] The aforementioned NiV-F protein, i) A 20-amino acid cleavage at or near the C-terminus of the wild-type NiV-F protein (SEQ ID NO: 2), and / or ii) Point mutations in N-linked glycosylation sites A targeted lipid particle of any of Invention 1001-1013 and 1022-1037, or a lentiviral vector of any of Invention 1014-1037, which includes the biologically active portion thereof. [Invention 1039] The NiV-F protein or the biologically active portion is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89%. Alternatively, a targeted lipid particle or lentiviral vector of the present invention 1038 having an amino acid sequence having sequence identity of approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%. [Invention 1040] The NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 22-amino acid cleavage at or near its C-terminus, wherein the targeted lipid particles or lentiviral vectors of any of Invention 1001-1013 and 1022-1037 are described in Invention 1014-1037. [Invention 1041] The NiV-F protein or the biologically active portion is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, and Targeted lipid particles or lentiviral vectors of the present invention 1040, having an amino acid sequence encoded by a sequence of nucleotides encoding a sequence having 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity. [Invention 1042] A targeted lipid particle according to any of the present inventions 1001-1013 and 1022-1041, or a lentiviral vector according to any of the present inventions 1014-1041, wherein the F protein comprises the sequence shown in SEQ ID NO: 23, and the G protein comprises the sequence shown in SEQ ID NO: 16. [Invention 1043] Targeted lipid particles according to any of the present invention 1001-1013 and 1022-1042, wherein the lipid bilayer is derived from the membrane of a host cell used to generate retroviruses or retrovirus-like particles. [Invention 1044] Targeted lipid particles of the present invention 1043, wherein the host cell is selected from the group consisting of CHO cells, BHK cells, MDCK cells, C3H 10T1 / 2 cells, FLY cells, Psi-2 cells, BOSC 23 cells, PA317 cells, WEHI cells, COS cells, BSC 1 cells, BSC 40 cells, BMT 10 cells, VERO cells, W138 cells, MRC5 cells, A549 cells, HT1080 cells, 293 cells, 293T cells, B-50 cells, 3T3 cells, NIH3T3 cells, HepG2 cells, Saos-2 cells, Huh7 cells, HeLa cells, W163 cells, 211 cells, and 211A cells. [Invention 1045] Targeted lipid particles according to any of the present invention 1001-1013 and 1022-1044, wherein the lipid bilayer is a viral envelope or contains one. [Invention 1046] Targeted lipid particles according to any of invention 1001-1013 and 1022-1045, comprising one or more viral components other than the F protein molecule and the G protein. [Invention 1047] Targeted lipid particles of the present invention 1046, wherein one or more viral components are derived from retroviruses. [Invention 1048] A targeted lipid particle according to any one of the present invention 1043 to 1047, wherein the retrovirus is a lentivirus or a lentivirus-like particle. [Invention 1049] A targeted lipid particle according to any of invention 1001-1013 and 1022-1048, or a lentiviral vector according to any of invention 1014-1042, further comprising an exogenous substance. [Invention 1050] The exogenous substance is present in the lumen of the targeted lipid particle or lentiviral vector according to the present invention 1049. [Invention 1051] A targeted lipid particle or lentiviral vector according to Invention 1049 or Invention 1050, wherein the exogenous substance is a protein or nucleic acid, and optionally the nucleic acid is DNA or RNA. [Invention 1052] A targeted lipid particle or lentiviral vector according to any one of the present invention 1049 to 1051, wherein the exogenous substance is a nucleic acid encoding cargo for delivery to the target cells. [Invention 1053] The exogenous substance is a targeted lipid particle or lentiviral vector according to any of items 1049 to 1052 of the present invention, wherein the exogenous substance encodes a therapeutic agent or a diagnostic agent. [Invention 1054] A targeted lipid particle or lentiviral vector according to any of items 1049 to 1053 of the present invention, wherein the exogenous substance encodes a membrane protein, and optionally the membrane protein is an antigen receptor for targeting cells expressed by or associated with a disease or pathological condition. [Invention 1055] The targeted lipid particle or lentiviral vector of the present invention 1053, wherein the membrane protein is a chimeric antigen receptor (CAR). [Invention 1056] A targeted lipid particle or lentiviral vector according to any of items 1052 to 1055 of the present invention, wherein the target cell is a T cell. [Invention 1057] The exogenous substance is a nucleic acid containing a gene defect, optionally a payload gene for correcting the gene defect in the target cell, and optionally the gene defect is a liver cell or a liver cell-related targeted lipid particle or lentiviral vector according to any of the present invention 1049 to 1053. [Invention 1058] A polynucleotide comprising a nucleic acid sequence encoding (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and (ii) a single-domain antibody (sdAb) variable domain, wherein the sdAb variable domain is conjugated to the C-terminus of the G protein or a biologically active portion thereof. [Invention 1059] The polynucleotide of the present invention 1058, wherein the single-domain antibody binds to cell surface molecules present on target cells. [Invention 1060] The polynucleotide of the present invention 1059, wherein the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule. [Invention 1061] The polynucleotide of Invention 1059 or Invention 1060, wherein the target cells are selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. [Invention 1062] The target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytic cells, SLC7A10+ adipocytes, or CD30+ lung epithelial cells, and are any of the polynucleotides according to invention 1059 to 1061. [Invention 1063] The cell surface molecule or antigen is selected from the group consisting of ASGR1, ASGR2, TM4SF5, CD8, CD4, and low-density lipoprotein receptor (LDL-R), and is a polynucleotide according to any of the invention 1059 to 1062. [Invention 1064] A polynucleotide comprising a nucleic acid sequence encoding (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and (ii) a binding domain that binds to a cell surface molecule selected from the group consisting of ASGR1, ASGR2, TM4SF5, CD4, CD8, and low-density lipoprotein receptor (LDL-R). [Invention 1065] The polynucleotide of the present invention 1064, wherein the binding domain is a single-domain antibody (sdAb) or a single-stranded variable fragment (scFv). [Invention 1066] A polynucleotide according to any one of the invention 1058 to 1065, wherein the nucleic acid sequence is a first nucleic acid sequence, and the polynucleotide further comprises a second nucleic acid sequence encoding a henipavirus F protein molecule or a biologically active portion thereof. [Invention 1067] The polynucleotide of the present invention 1066, wherein the polynucleotide comprises an IRES or a sequence encoding a linking peptide between the first and second nucleic acid sequences, and optionally the linking peptide is a self-cleaving peptide or a peptide that causes ribosome skipping, optionally a T2A peptide. [Invention 1068] A polynucleotide according to any of the present invention 1058 to 1067, further comprising the expression of the nucleic acid, and at least one promoter functionally linked to control the expression of the first nucleic acid sequence and the second nucleic acid sequence. [Invention 1069] A polynucleotide according to any one of the present invention 1058 to 1068, wherein the sdAb variable domain or the binding domain is bound to the G protein via an encoded peptide linker. [Invention 1070] The polynucleotide of the present invention 1069, wherein the encoded peptide linker has a length of up to 65 amino acids, and optionally between 2 and 65 amino acids. [Invention 1071] The polynucleotide of Invention 1069 or Invention 1070, wherein the encoded peptide linker comprises GS, GGS, GGGGS (SEQ ID NO: 43), GGGGGS (SEQ ID NO: 41) and combinations thereof, or the encoded peptide linker comprises (GGS)n where n is 1 to 10, (GGGGS)n where n is 1 to 10 (SEQ ID NO: 42), or (GGGGGS)n where n is 1 to 4 (SEQ ID NO: 27). [Invention 1072] The polynucleotide of any of the Invention 1058-1071, wherein the encoded G protein is wild-type Nipah virus G (NiV-G) protein or Hendra virus G protein, or a functionally active variant or biologically active moiety thereof, wherein the variant is a variant that exhibits reduced binding to its native binding partner. [Invention 1073] The polynucleotide according to any of items 1058 to 1072 of the present invention, wherein the encoded G protein is a wild-type NiV-G protein or a functionally active variant or a biologically active moiety thereof. [Invention 1074] The NiV-G protein or its functionally active variant or biologically active moiety comprises the amino acid sequence shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, or comprises at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, and A polynucleotide according to Invention 1072 or Invention 1073, comprising an amino acid sequence having sequence identity of 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1075] The aforementioned NiV-G protein, A cleaved and biologically active moiety lacking up to 40 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44). The polynucleotides are any of the 1072-1074 of the present invention. [Invention 1076] The NiV-G protein is the biologically active portion cleaved at the N-terminus of wild-type NiV-G, and is at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, less than A polynucleotide according to any of Invention 1072 to 1075, comprising an amino acid sequence having sequence identity of 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1077] The NiV-G protein is a biologically active moiety having a cleavage at or near the N-terminus of the wild-type NiV-G protein, selected from the group consisting of a 5-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 10-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 15-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 20-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 25-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, a 30-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, or a 34-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein, wherein the wild-type NiV-G protein is optionally a polynucleotide of any of the present inventions 1072 to 1076 as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44. [Invention 1078] The polynucleotide of any of the present invention 1072 to 1077, wherein the NiV-G protein is a biologically active portion having a 34-amino acid cleavage at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44). [Invention 1079] The polynucleotide according to any of the invention 1058 to 1078, wherein the G protein is a mutant NiV-G protein that exhibits reduced binding to ephrin B2 or ephrin B3. [Invention 1080] The polynucleotide of the present invention 1079, wherein the mutant NiV-G protein contains one or more amino acid substitutions corresponding to amino acid substitutions selected from the group consisting of E501A, W504A, Q530A and E533A with reference to the numbering shown in Sequence ID No. 28. [Invention 1081] The polynucleotide of Invention 1079 or Invention 1080, comprising the mutant NiV-G protein, with amino acid substitutions E501A, W504A, Q530A, and E533A, with reference to the numbering shown in Sequence ID No. 28. [Invention 1082] The polynucleotide according to any of the Invention 1079 to 1081, wherein the mutant NiV-G protein comprises an amino acid sequence having at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 16. [Invention 1083] The polynucleotide according to any of items 1066 to 1082 of the present invention, wherein the F protein or its biologically active portion is wild-type Nipah virus F (NiV-F) protein or Hendra virus F protein, or a functionally active variant or biologically active portion thereof. [Invention 1084] A polynucleotide according to any of the present invention 1066 to 1083, wherein the F protein or its biologically active portion is a wild-type NiV-F protein or a functionally active variant or biologically active portion. [Invention 1085] The NiV-F protein or its functionally active variant or biologically active moiety is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least A polynucleotide according to Invention 1083 or Invention 1084, comprising an amino acid sequence having sequence identity of 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1086] The aforementioned NiV-F protein, i) A 20-amino acid cleavage at or near the C-terminus of the wild-type NiV-F protein (SEQ ID NO: 2), and / or ii) Point mutations in N-linked glycosylation sites The polynucleotide of Invention 1085 or Invention 1085, which includes the biologically active portion thereof. [Invention 1087] The NiV-F protein or the biologically active portion is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or less A polynucleotide of the present invention 1086 having an amino acid sequence having sequence identity of at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. [Invention 1088] The polynucleotide of Invention 1085 or Invention 1086, wherein the NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 22-amino acid cleavage at or near its C-terminus. [Invention 1089] The NiV-F protein or the biologically active portion is 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 8 A polynucleotide of the present invention 1088, having an amino acid sequence encoded by a sequence of nucleotides encoding a sequence having 9%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity. [Invention 1090] A polynucleotide according to any of the present invention 1066 to 1089, wherein the F protein contains the sequence shown in SEQ ID NO: 23, and the G protein contains the sequence shown in SEQ ID NO: 16. [Invention 1091] A vector comprising any of the polynucleotides 1058 to 1090 of the present invention. [Invention 1092] The vector of the present invention 1091, wherein the vector is a mammalian vector, a viral vector, or an artificial chromosome, and optionally the artificial chromosome is a bacterial artificial chromosome (BAC). [Invention 1093] A plasmid comprising any of the polynucleotides 1058 to 1090 of the present invention. [Invention 1094] A plasmid of the present invention 1093, further comprising one or more nucleic acids encoding proteins for lentivirus generation. [Invention 1095] A cell comprising any polynucleotide of Invention 1058 to 1090, or a vector of Invention 1091 or Invention 1092, or a plasmid of Invention 1093 or Invention 1094. [Invention 1096] A method for producing targeted lipid particles comprising a henipavirus F protein molecule or its biologically active portion, and a targeted envelope protein comprising a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and a single-domain antibody (sdAb) variable domain, a) To provide a cell comprising a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and a single-domain antibody (sdAb) variable domain, b) Culturing the cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying the targeted lipid particles from the cells, thereby producing the targeted lipid particles. The method, including the method described above. [Invention 1097] A method for producing a pseudotyped lentiviral vector, a) To provide producer cells comprising lentiviral nucleic acids (multiple) and nucleic acids encoding a henipavirus F protein molecule or a biologically active portion thereof and nucleic acids encoding a targeted envelope protein, wherein the targeted envelope protein comprises henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and a single-domain antibody. b) Culturing the cells under conditions that enable the generation of the lentiviral vector, and c) Isolating, concentrating, or purifying the lentiviral vector from the cells, thereby producing the pseudotyped lentiviral vector. The method, including the method described above. [Invention 1098] A method for producing targeted lipid particles comprising a henipavirus F protein molecule or its biologically active portion, and a targeted envelope protein comprising a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and binding domain, a) To provide a cell comprising a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion and a binding domain, and the binding domain is (i) ASGR1, ASGR2, and TM4SF5, or any cell surface molecule selected from the group consisting of human ASGR1, human ASGR2, and human ASGR2, (ii) A cell surface molecule that binds to CD4 or CD8, or is selected from the group consisting of human CD4 or human CD8, or (iii) Low-density lipoprotein receptor (LDL-R), which optionally binds to a cell surface molecule that is human LDL-R. To provide as described above, b) Culturing the cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying the targeted lipid particles from the cells, thereby producing the targeted lipid particles. The method, including the method described above. [Invention 1099] A method for producing a pseudotyped lentiviral vector, a) Providing producer cells comprising lentiviral nucleic acid(s), a nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and a nucleic acid encoding a targeted envelope protein, wherein the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion and a binding domain, and the binding domain is (i) ASGR1, ASGR2, and TM4SF5, or any cell surface molecule selected from the group consisting of human ASGR1, human ASGR2, and human ASGR2, (ii) A cell surface molecule that binds to CD4 or CD8, or is selected from the group consisting of human CD4 or human CD8, or (iii) Low-density lipoprotein receptor (LDL-R), which optionally binds to a cell surface molecule that is human LDL-R. To provide as described above, b) Culturing the producer cells under conditions that enable the generation of lentiviral vectors, and c) Isolating, concentrating, or purifying the lentiviral vector from the cells, thereby producing the pseudotyped lentiviral vector. The method, including the method described above. [Invention 1100] The method of Invention 1098 or Invention 1099, wherein the binding domain is a single-domain antibody or a single-stranded variable fragment (scFv). [Invention 1101] A method for producing targeted lipid particles comprising a henipavirus F protein molecule or its biologically active portion and a targeted envelope protein, a) To provide cells containing any polynucleotide of Invention 1058 to 1090, or a vector of Invention 1091 or Invention 1092, or a plasmid of Invention 1093 or Invention 1094, b) Culturing the cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying the targeted lipid particles from the cells, thereby producing the targeted lipid particles. The method, including the method described above. [Invention 1102] A method for producing a pseudotyped lentiviral vector, a) To provide producer cells comprising lentiviral nucleic acid(s) and any polynucleotide of Invention 1058-1090, or a vector of Invention 1091 or Invention 1092, or a plasmid of Invention 1093 or Invention 1094. b) Culturing the cells under conditions that enable the generation of the lentiviral vector, and c) Isolating, concentrating, or purifying the lentiviral vector from the cells, thereby producing the pseudotyped lentiviral vector. The method, including the method described above. [Invention 1103] The method of the present invention 1101 or 1102 further comprises providing the cells with a henipavirus F protein molecule or a polynucleotide encoding a biologically active portion thereof, prior to step (b). [Invention 1104] The method according to any one of the present invention 1096 to 1103, wherein the cells are mammalian cells. [Invention 1105] The method according to any one of the present invention 1096 to 1104, wherein the cells are producer cells containing viral nucleic acid, optionally retroviral nucleic acid, or lentiviral nucleic acid, and the targeted lipid particles are viral particles or virus-like particles, optionally retroviral particles or retroviral-like particles, optionally lentiviral particles or lentiviral-like particles. [Invention 1106] Producer cells comprising any polynucleotide of Invention 1058 to 1090, or a vector of Invention 1091 or Invention 1092, or a plasmid of Invention 1093 or Invention 1094. [Invention 1107] Producer cells of the present invention 1106, further comprising a nucleic acid encoding the henipavirus F protein or a biologically active portion thereof. [Invention 1108] The producer cell of the present invention 1106 or 1107, wherein the cell further contains viral nucleic acid, and optionally the viral nucleic acid is lentiviral nucleic acid. [Invention 1109] A producer cell comprising (i) viral nucleic acid(s), (ii) nucleic acid encoding a henipavirus F protein molecule or a biologically active portion thereof, and (iii) nucleic acid encoding a targeted envelope protein including henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion thereof and a single-domain antibody (sdAb) variable domain, wherein the viral nucleic acid(s) is optionally lentiviral nucleic acid. [Invention 1110] The producer cell of the present invention 1109, wherein the single-domain antibody binds to cell surface molecules present on the target cell. [Invention 1111] The producer cell of the present invention 1110, wherein the cell surface molecule is a protein, glycan, lipid, or low molecular weight molecule. [Invention 1112] Producer cells of the present invention 1110 or 1111, wherein the target cells are selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. [Invention 1113] Producer cells according to any of the present invention 1110 to 1112, wherein the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytic cells, SLC7A10+ adipocytes, or CD30+ lung epithelial cells. [Invention 1114] A producer cell comprising (i) viral nucleic acids(s), (ii) nucleic acids encoding a henipavirus F protein molecule or a biologically active portion thereof, and (iii) nucleic acids encoding a targeted envelope protein including a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active portion and a binding domain, wherein the binding domain is (i) ASGR1, ASGR2, and TM4SF5, or any cell surface molecule selected from the group consisting of human ASGR1, human ASGR2, and human ASGR2, (ii) A cell surface molecule that binds to CD4 or CD8, or is selected from the group consisting of human CD4 or human CD8, or (iii) Low-density lipoprotein receptor (LDL-R), optionally binding to a cell surface molecule which is human LDL-R, The viral nucleic acid(s) may be any one of the above, and the viral nucleic acid(s) may be a lentiviral nucleic acid. The aforementioned producer cells. [Invention 1115] Producer cells of the present invention 1114, wherein the binding domain is a single-domain antibody or a single-stranded variable fragment. [Invention 1116] Producer cells according to any of the inventions 1109 to 1115, wherein the F protein or its biologically active portion is wild-type Nipah virus F (NiV-F) protein or Hendra virus F protein, or a functionally active variant or biologically active portion thereof. [Invention 1117] A producer of any of the inventions 1109 to 1116, wherein the F protein or its biologically active portion is a wild-type NiV-F protein or a functionally active variant or biologically active portion thereof. [Invention 1118] The henipavirus F protein molecule or its biologically active portion (i) Sequences shown in Sequence ID 2, Sequence ID 5, Sequence ID 7, Sequence ID 8 or Sequence ID 23, (ii) Amino acid sequences having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 23. Producer cells according to any of the inventions 1109 to 1117, including the above. [Invention 1119] Producer cells according to any of the inventions 1109 to 1118, wherein the encoded G protein is wild-type Nipah virus G (NiV-G) protein or Hendra virus G protein, or a functionally active variant or biologically active moiety thereof, wherein the variant is optionally a variant that exhibits reduced binding to its native binding partner. [Invention 1120] Producer cells according to any of invention 1109 to 1119, wherein the encoded G protein is wild-type NiV-G protein or a functionally active variant or biologically active portion thereof. [Invention 1121] The henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion (i) Sequence shown in Sequence ID 9, Sequence ID 28, or Sequence ID 44, (ii) Amino acid sequences having 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44 Producer cells, including any of the present inventions 1109 to 1120. [Invention 1122] The henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion (i) Sequences shown in sequence numbers 10-16, 35-40 or 45-51, (ii) Amino acid sequences having sequence identity of at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. Producer cells according to any of the inventions 1109 to 1121, including the above. [Invention 1123] The producer cells have membrane (e.g., cell membrane) expression of the targeting envelope protein, which is more abundant in the membrane (e.g., cell membrane) compared to reference producer cells in which the same envelope protein is incorporated into the membrane (e.g., cell membrane) but is fused to the alternative targeting site, and optionally the alternative targeting site is a single-stranded variable fragment (scFv), and / or The producer cells express the targeted envelope protein on the membrane (e.g., cell membrane) of the producer cells, which is at least 20 proteins per square micron (e.g., at least 50, 100, 200, 500, 1000, 2000, 5000, or 10,000 proteins), and / or the targeted envelope protein constitutes at least 0.1% (e.g., at least 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%) (e.g., relative to the total protein weight) of the producer cells' total membrane (e.g., cell membrane) proteins. Producer cells according to any of inventions 1109 to 1122. [Invention 1124] Targeted lipid particles or pseudotyped lentiviral vectors generated by any of the methods 1096 to 1105 of the present invention or from producer cells according to any of the methods 1106 to 1122 of the present invention. [Invention 1125] The targeted lipid particle according to any of the present invention 1001-1013, 1021-1057, and 1124, wherein the targeted lipid particle has a higher expression of the targeted envelope protein compared to a reference lipid particle in which the same envelope protein is incorporated into a similar lipid bilayer but is fused to an alternative targeting site, and optionally the alternative targeting site is a single-stranded variable fragment (scFv). [Invention 1126] The titer in target cells after transduction is 1 × 10⁻⁶ 6 Transduction units (TU) / mL or more, 2 × 10 6 TU / mL or more, 3×10 6 TU / mL or more, 4×10 6 TU / mL or more, 5×10 6 TU / mL or more, 6×10 6 TU / mL or more, 7×10 6 TU / mL or more, 8×10 6 TU / mL or more, 9x10 6 TU / mL or higher, or 1x10 7 TU / mL or higher, and / or The targeting envelope protein is at least approximately (0.001, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, or 0.5) in size per nm. 2 The density present on the surface of the targeted lipid particles is Targeted lipid particles according to any of invention 1001-1013, 1021-1057, 1124 and 1125, or lentiviral vectors according to any of invention 1013-1042, 1049-1057 and 1124. [Invention 1127] A composition comprising any of the multiple targeted lipid particles or multiple lentiviral vectors according to invention 1001-1057 and 1124-1126. [Invention 1128] The composition of the present invention 1127, further comprising a pharmaceutically acceptable carrier. [Invention 1129] In the population of lipid particles or lentiviral vectors in the composition, 50% or more than 50%, 55% or more than 55%, 60% or more than 60%, 65% or more than 65%, 70% or more than 70%, or 75% or more than 75% are surface positive for the targeted envelope protein, and / or The targeting envelope protein is at least approximately (0.001, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2, or 0.5) in size per nm. 2 The average density present on the surface of the targeted lipid particles is A composition according to Invention 1127 or Invention 1128. [Invention 1130] A method for transducing cells, comprising transducing cells with a lentiviral vector according to any of the present invention 1013-1042, 1049-1057 and 1124, or a composition comprising any of the lentiviral vectors according to the present invention 1013-1042, 1049-1057 and 1124, or a plurality of lentiviral vectors. [Invention 1131] The targeted envelope protein of the lentiviral vector or targeted lipid particle targets CD4, and the cell is a CD4+ cell, or The targeted envelope protein of the lentiviral vector targets CD8, and the cell is a CD8+ cell, or The targeted envelope protein of the lentiviral vector targets ASGR1, ASGR2, or TM4SF5, and the cells are hepatocytes. The method of the present invention 1130. [Invention 1132] A method for delivering an exogenous substance to a subject (e.g., a human subject), comprising administering to the subject any of the targeted lipid particles or lentiviral vectors according to invention 1049-1057, 1125, and 1126, wherein the targeted lipid particles or lentiviral vector contain the exogenous substance. [Invention 1133] A method for delivering an exogenous substance to a subject (e.g., a human subject), comprising administering to the subject any composition according to 1127 to 1129 of the present invention, wherein a plurality of targeted lipid particles or lentiviral vectors contain the exogenous substance. [Invention 1134] A method for delivering a chimeric antigen receptor (CAR) to a cell, comprising contacting the cell with a lentiviral vector according to any of the inventions 1014-1016 and 1019-1042 or a targeted lipid particle according to the invention 1055 or the invention 1056, wherein the lentiviral vector or targeted lipid particle contains a nucleic acid encoding the CAR. [Invention 1135] A method for delivering a chimeric antigen receptor (CAR) to a cell, comprising contacting the cell with any composition from 1127 to 1129 of the present invention, wherein a plurality of lentiviral vectors or targeted lipid particles comprise a nucleic acid encoding the CAR. [Invention 1136] A method for delivering an exogenous substance to hepatocytes, comprising contacting the cells with a lentiviral vector according to any of the inventions 1017 to 1042 or with targeted lipid particles or a lentiviral vector according to the invention 1057. [Invention 1137] A method for delivering an exogenous substance to hepatocytes, comprising contacting the cells with any of the compositions of the present invention 1127 to 1129, wherein a plurality of lentiviral vectors or targeted lipid particles contain the exogenous substance for delivery to the hepatocytes. [Invention 1138] Any method 1134 to 1137 of the present invention, wherein the contact comprises transduction into the cells with a lentiviral vector or the targeted lipid particles, and optionally the contact is in vivo in the subject. [Invention 1139] A method for treating a disease or disorder in a subject (e.g., a human subject), comprising administering to the subject any of the targeted lipid particles of the present invention 1001-1013, 1021-1057, and 1124-1126, or any lentiviral vector of the present invention 1013-1042, 1049-1057, 1124, and 1126, or any of the compositions of the present invention 1127-1129. [Invention 1140] A method for fusing mammalian cells to targeted lipid particles, comprising administering to a subject any of the targeted lipid particles according to any of the present inventions 1001-1013, 1021-1057, and 1124-1126, a lentiviral vector according to any of the present inventions 1013-1042, 1049-1057, 1124, and 1126, or any of the compositions according to the present inventions 1127-1129. [Invention 1141] The method of the present invention 1140, wherein fusing the mammalian cells with the targeted lipid particles delivers an exogenous substance to a target (for example, a human target). [Brief explanation of the drawing]

[0157] [Figure 1A] Figures 1A–1C illustrate the characterization of cells transfected with constructs containing scFv or VHH-binding modalities. Figure 1A shows the surface expression of cells transfected with constructs containing scFv or VHH-binding modalities, shown as median fluorescence intensity (MFI) quantified by the percentage of His+ cells, analyzed by flow cytometry. Empty vectors and expression vectors without binder domains were used as negative controls. [Figure 1B] Figures 1A–1C show the characterization of cells transfected with constructs containing scFv or VHH-binding modality. Figure 1B shows the binding of cells transfected with constructs containing the VHH-binding modality scFV to the soluble hCD4-Fc protein, as analyzed by flow cytometry and shown as median fluorescence intensity (MFI) quantified by %Fc+ cells. Empty vectors and expression vectors without a binder domain were used as negative controls. [Figure 1C]Figures 1A–1C illustrate the characterization of cells transfected with constructs containing scFv or VHH binding modalities. Figure 1C shows the surface expression of targeted binding sequences on 293 cells transfected with constructs containing VHH binding modalities, compared to the scFv binding modality, as analyzed by flow cytometry and expressed as median fluorescence intensity (MFI), quantified by the percentage of His+ cells. Empty vectors and expression vectors without binder domains were used as negative controls. [Figure 2] We demonstrate the transduction efficacy of four exemplary constructs containing scFV or VHH-binding modalities to PanT cells from peripheral blood that was negatively selected, thawed, and activated with anti-CD3 / anti-CD28 to enrich T cells. Cells were analyzed by flow cytometry, and titer was determined by the percentage of CD4-positive cells that were GFP+. [Figure 3A] Figures 3A-B show the transduction efficiency of CD8 retargeting pseudotyped lentivirus in an in vivo model using activated PBMCs intraperitoneally injected into NOD-scid-IL2rγnull mice, as analyzed by flow cytometry. Transduction efficiency of CD8 retargeting pseudotyped lentivirus is shown for CD8+ (Figure 3A) or CD8- (Figure 3B) T cells, and titers were determined by the percentage of CD8-positive or CD8-negative cells that were GFP+. [Figure 3B] See the explanation in Figure 3A. [Figure 4A] Figures 4A and 4B show the ability of CD8 retargeting pseudotyped lentiviruses containing chimeric antigen receptors (CARs) to kill leukemia cells in vitro. Figure 4A shows the ability to detect CD19+CAR expression on CD8+ cells 4 days after transduction. Figure 4B shows the elimination of Nalm6 cells, assessed at 18 hours after incubation, as analyzed by flow cytometry. [Figure 4B] See the explanation in Figure 4A. [Modes for carrying out the invention]

[0158] I. Definition Unless otherwise defined, all terms, notations, and other technical and scientific or specialized terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art in which the claimed subject matter relates. In some cases, terms having a commonly understood meaning are defined herein for clarity and / or for immediate reference, and the inclusion of such definitions herein should not necessarily be construed as indicating a substantial difference from the commonly understood meaning in the art.

[0159] Unless otherwise defined, all technical and scientific terms, acronyms, and abbreviations used herein have the same meaning as those generally understood by those skilled in the art in the field to which this invention relates. Unless otherwise indicated, abbreviations and symbols for chemical and biochemical names follow IUPAC-IUB nomenclature. Unless otherwise indicated, all numerical ranges include the values ​​defining the range and all integer values ​​between them.

[0160] As used herein, the articles "a" and "an" refer to one or more (i.e., at least one) grammatical objects of the article. For example, "an element" means one or more elements.

[0161] As used herein, “about” is to be understood by those skilled in the art and varies to some extent in the context in which it is used. As used herein, when “about” refers to a measurable value, such as a quantity, duration of time, etc., it includes a variation of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and more preferably ±0.1% from the specified value, as such variation is appropriate for carrying out the disclosed method.

[0162] As used herein, “lipid particle” refers to any biological or synthetic particle containing a bilayer of amphiphilic lipids surrounding its lumen or cavity. Typically, lipid particles do not contain a nucleus. Examples of lipid particles include solid particles, e.g., nanoparticles, virus-derived particles, or cell-derived particles. Such lipid particles include, but are not limited to, virus particles (e.g., lentiviral particles), virus-like particles, viral vectors (e.g., lentiviral vectors), exosomes, enucleated cells, various vesicles, e.g., microvesicles, membrane vesicles, extracellular membrane vesicles, cell membrane vesicles, giant cell membrane vesicles, apoptotic bodies, mitoparticles, pyrenocytes, or lysosomes. In some embodiments, the lipid particle may be a fusosome. In some embodiments, the lipid particle is not a platelet.

[0163] As used herein, with respect to proteins such as G proteins or F proteins, “biologically active moiety” refers to a portion of a protein that exhibits or retains the activity or properties of the full-length protein. For example, the biologically active moiety of an F protein, together with a G protein, retains fusion activity when each is embedded in a lipid bilayer. The biologically active moiety of a G protein, together with an F protein, retains fusion activity when each is embedded in a lipid bilayer. The retained activity may range from 10% to 150% or more of the activity of the full-length or wild-type F protein or G protein. Examples of biologically active regions of F and G proteins include cleavage of the cytoplasmic domain, e.g., cleavage of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35 or more consecutive amino acids; see, for example, Khetawat and Broder 2010 Virology Journal 7:312, Witting et al. 2013 Gene Therapy 20:997-1005, and patent application no. WO / 2013 / 148327.

[0164] As used herein, “fusosome” refers to a particle containing an amphiphilic lipid bilayer surrounding a lumen or cavity and a fusogen that interacts with the amphiphilic lipid bilayer. In embodiments, fusosomes contain nucleic acids. In some embodiments, fusosomes are membrane-enveloped preparations. In some embodiments, fusosomes are derived from source cells.

[0165] As used herein, “fusosome composition” refers to a composition comprising one or more fusosomes.

[0166] As used herein, "fusogen" refers to a substance or molecule that creates an interaction between two membrane-enveloping lumens. In some embodiments, the fusogen facilitates membrane fusion. In other embodiments, the fusogen creates a connection, e.g., a pore, between two lumens (e.g., the lumen of a retroviral vector and the cytoplasm of a target cell). In some embodiments, the fusogen comprises a complex of two or more proteins, e.g., none of the proteins possess fusion activity individually. In some embodiments, the fusogen comprises a targeting domain.

[0167] As used herein, “retargeted fusogen” refers to a fusogen that includes a targeting site having a sequence that is not part of the naturally occurring form of the fusogen. In embodiments, the fusogen includes a different targeting site compared to the targeting site in the naturally occurring form of the fusogen. In embodiments, the naturally occurring form of the fusogen lacks a retargeting main, and the retargeted fusogen includes a targeting site that is not present in the naturally occurring form of the fusogen. In embodiments, the fusogen is modified to include a targeting site. In embodiments, the fusogen includes, for example, one or more sequence modifications outside the targeting site in a transmembrane domain, a fusionally active domain, or a cytoplasmic domain compared to the naturally occurring form of the fusogen.

[0168] As used herein, “targeted envelope protein” refers to a polypeptide containing a single-domain antibody (sdAb) variable domain that targets a molecule to a desired cell type, e.g., a VL or VH-only sdAb, a nanobody, a camelid VHH domain, a shark IgNAR, or a henipavirus G protein conjugated to a fragment thereof. In some such embodiments, conjugation may be direct or indirect, via a linker such as a peptide linker.

[0169] As used herein, “targeted lipid particles” refers to lipid particles containing targeted envelope proteins embedded in a lipid bilayer.

[0170] As used herein, “retroviral nucleic acid” refers to a nucleic acid containing at least the minimum sequence requirements for packaging a retrovirus or retroviral vector, either alone or in combination with a helper cell, helper virus, or helper plasmid. In some embodiments, the retroviral nucleic acid further comprises or encodes an exogenous substance, a positive target cell-specific regulatory element, a non-target cell-specific regulatory element, or a negative TCSRE. In some embodiments, the retroviral nucleic acid comprises one or more (e.g., all) of the following: 5'LTR (e.g., for promoting integration), U3 (e.g., for activating viral genomic RNA transcription), R (e.g., a Tat-binding region), U5, 3'LTR (e.g., for promoting integration), a packaging site (e.g., psi(Ψ)), and RRE (e.g., for binding to Rev and promoting nuclear export). The retroviral nucleic acid may comprise RNA (e.g., if it is part of a virion) or DNA (e.g., if it has been introduced into a source cell or after reverse transcription in a recipient cell). In some embodiments, retroviral nucleic acids are packaged using helper cells, helper viruses, or helper plasmids containing one or more (e.g., all) of gag, pol, and env.

[0171] As used herein, “target cells” refers to the type of cells to which targeted lipid particles are desired to deliver exogenous substances. In embodiments, target cells are cells of a specific tissue type or class, e.g., immune effector cells, e.g., T cells. In some embodiments, target cells are diseased cells, e.g., cancer cells. In some embodiments, fusogens, e.g., retargeted fusogens, result in preferential delivery of exogenous substances to target cells compared to non-target cells.

[0172] As used herein, “non-target cells” refers to a type of cell to which targeted lipid particles do not wish to deliver exogenous substances. In some embodiments, non-target cells are cells of a particular tissue type or class. In some embodiments, non-target cells are non-affected cells, e.g., non-cancer cells. In some embodiments, fusogens, e.g., retargeted fusogens, result in lower delivery of exogenous substances to non-target cells compared to target cells.

[0173] As used herein, “single-domain antibody” or “sdAb” refers to an antibody having a single monomeric domain antigen-binding / recognition domain. Such antibodies include nanobodies, camelid antibodies (e.g., VHH), or shark antibodies (e.g., IgNAR). In some embodiments, the variable domain of the sdAb comprises three CDRs and four framework regions (designated as FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4). In some embodiments, the sdAb variable domain may be cleaved at the N-terminus or C-terminus to contain only partial FR1 and / or FR4, or to lack one or both of those framework regions, as long as the sdAb variable domain substantially maintains antigen-binding and specificity.

[0174] The term "CDR" refers to a complementarity determination region defined by at least one identification method for those skilled in the art. The precise amino acid sequence boundaries of a given CDR or FR are as follows: Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 (“Chothia” numbering scheme); MacCallum et al., J.Mol.Biol.262:732-745 (1996), “Antibody-antigen interactions: Contact analysis and binding site topography,” J.Mol.Biol.262, 732-745.” (“Contact” numbering scheme); Lefranc MP et al., “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains,” Dev Comp Immunol, 2003. The numbering scheme can be easily determined using one of several well-known schemes, including those described in Jan;27(1):55-77 ("IMGT" numbering scheme), Honegger A and Pluckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8;309(3):657-70, ("Aho" numbering scheme), and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86(23):9268-9272, ("AbM" numbering scheme).

[0175] The boundaries of a given CDR or FR can vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. For both the Kabat and Chothia schemes, numbering is based on the most common antibody region sequence length, with insertions provided by insertive letters, e.g., "30a", and deletions that appear in some antibodies. These two schemes place given insertions and deletions ("indels") at different positions, resulting in different numbering. The Contact scheme is based on the analysis of complex crystal structures and is similar in many ways to the Chothia numbering scheme. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on those used by Oxford Molecular's AbM antibody modeling software.

[0176] In some embodiments, CDRs may be defined according to one of the following: the Chothia numbering scheme, the Kabat numbering scheme, a combination of Kabat and Chothia, the AbM definition, and / or the Contact definition. The sdAb variable domain includes three CDRs designated as CDR1, CDR2, and CDR3. Table 1 below lists exemplary positional boundaries of CDR-H1, CDR-H2, and CDR-H3 as identified by the Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, residue numbering is listed using both the Kabat and Chothia numbering schemes. FRs are located between CDRs; for example, FR-H1 is located before CDR-H1, FR-H2 is located between CDR-H1 and CDR-H2, FR-H3 is located between CDR-H2 and CDR-H3, and so on. Note that the Kabat numbering scheme shown places insertions at H35A and H35B, so the ends of a Chothia CDR-H1 loop will vary between H32 and H34 depending on the loop length when numbered using the Kabat numbering rules shown.

[0177] (Table 1) Boundaries of CDRs following various numbering schemes TIFF0007887223000001.tif77165

[0178] Therefore, unless otherwise specified, a given antibody or its variable region, or any "CDR" or "complementarity-determining region" in that region, or any individual specific CDR (e.g., CDR-H1, CDR-H2, CDR-H3), should be understood to encompass a certain (or specific) complementarity-determining region as defined by any of the schemes described above. For example, if it is described that a particular CDR (e.g., CDR-H3) contains the amino acid sequence of the corresponding CDR in a given sdAb amino acid sequence, it is understood that such a CDR has the sequence of the corresponding CDR (e.g., CDR-H3) in the sdAb as defined by any of the schemes described above. It is understood that any antibody, e.g., an sdAb, contains a CDR, which can be identified according to any of the other numbering schemes described above or any other numbering scheme known to those skilled in the art.

[0179] As used herein, the term “specifically binds” to a target molecule such as an antigen means that the binding molecule, e.g., a single-domain antibody, reacts or associates with a particular target molecule more frequently, more rapidly, for a longer duration, and / or with a higher affinity than alternative molecules. A binding molecule, e.g., an sdAb variable domain, “specifically binds” to a target molecule if it binds with higher affinity, avidity, more readily, and / or for a longer duration than it would if it bound to other molecules. It is understood that a binding molecule such as an sdAb that specifically binds to a first target may or may not specifically bind to a second target. Thus, “specific binding” does not necessarily require (but may include) exclusive binding.

[0180] As used herein, with respect to peptide, polypeptide, or antibody sequences, “amino acid sequence identity percentage (%)” and “homology” are defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence, after the sequences have been aligned and gaps introduced as necessary to achieve the maximum sequence identity percentage, without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining amino acid sequence identity percentage can be achieved in various ways within the scope of the art, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN™ (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm necessary to achieve the maximum alignment over the entire length of the sequences being compared.

[0181] Amino acid substitutions may include, but are not limited to, the replacement of one amino acid with another in a polypeptide. Exemplary substitutions are shown in Table 2. Amino acid substitutions are introduced into the antibody of interest, and the product can be screened for desired activity, e.g., retention / improved binding.

[0182] (Table 2) TIFF0007887223000002.tif131165

[0183] Amino acids can be classified according to their common side-chain properties: (1) Hydrophobic: norleucine, Met, Ala, Val, Leu, Ile (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln, (3) Acidic: Asp, Glu, (4) Basicity: His, Lys, Arg, (5) Residues that affect chain orientation: Gly, Pro, (6) Aromatic: Trp, Tyr, Phe.

[0184] Non-conservative substitution involves swapping one member of one of these classes with one of another.

[0185] The term "corresponds" to a protein location, for example, the statement that a nucleotide or amino acid location "corresponds" to a nucleotide or amino acid location in a disclosed sequence, such as those shown in a sequence listing, refers to a nucleotide or amino acid location identified by alignment with the disclosed sequence based on structural sequence alignment or using a standard alignment algorithm such as the GAP algorithm. For example, corresponding residues in similar sequences (e.g., fragments or species variants) can be determined by alignment with a reference sequence by a structural alignment method. By aligning sequences, a person skilled in the art can identify corresponding residues, for example, using conserved and identical amino acid residues as guides.

[0186] The term “isolated,” as used herein, refers to a molecule that has been separated from at least a portion of its naturally occurring or typically produced components. For example, a polypeptide is referred to as “isolated” if it has been separated from at least a portion of the components of the cell in which it was produced. If a polypeptide is secreted by a cell after expression, physically separating the supernatant containing the polypeptide from the cell in which it was produced is considered “isolation” of the polypeptide. Similarly, a polynucleotide is referred to as “isolated” if it is not part of a larger polynucleotide that is naturally occurring or typically produced (e.g., in the case of a DNA polynucleotide, such as genomic DNA or mitochondrial DNA), or, for example, in the case of an RNA polynucleotide, if it has been separated from at least a portion of the components of the cell in which it was produced. Thus, a DNA polynucleotide contained in a vector inside a host cell may be referred to as “isolated.”

[0187] Where used herein, the term "effective dose" means an amount of a pharmaceutical composition sufficient to significantly and favorably modify (e.g., provide a favorable clinical response) the symptom and / or condition being treated. The effective dose of the active ingredient for use in a pharmaceutical composition varies with the knowledge and expertise of the physician, depending on the specific condition being treated, the severity of the condition, the duration of treatment, the characteristics of the combination therapy, the specific active ingredient(s) used, the specific pharmaceutically acceptable excipient(s) and / or carrier(s) used, and similar factors.

[0188] When used herein in relation to targeted lipid particles, “exogenous substance” refers to a substance that is not contained in, and does not encode, the corresponding wild-type virus or fusogen produced from the corresponding wild-type source cell. In some embodiments, the exogenous substance is a protein or nucleic acid that does not exist naturally and has a sequence modified (e.g., by insertion, addition, or substitution) relative to a naturally occurring protein. In some embodiments, the exogenous substance does not exist naturally in the source cell. In some embodiments, the exogenous substance is naturally present in the source cell but is exogenous with respect to the virus. In some embodiments, the exogenous substance does not exist naturally in the recipient cell. In some embodiments, the exogenous substance is naturally present in the recipient cell but is not present at a desired level or for a desired time. In some embodiments, the exogenous substance includes RNA or a protein.

[0189] As used herein, “promoter” refers to a cis-acting DNA sequence that, when functionally ligated to a gene coding sequence, induces the transcription of a gene. A promoter may include a transcription factor binding site. In some embodiments, a promoter acts in cooperation with one or more enhancers distal to the gene.

[0190] As used herein, composition refers to any mixture of two or more products, substances, or compounds, including cells. It may be a solution, suspension, liquid, powder, paste, aqueous, non-aqueous, or any combination thereof.

[0191] As used herein, the term “pharmaceutically acceptable” means a substance such as a carrier or diluent that does not inhibit the biological activity or properties of a compound and is relatively non-toxic; that is, the substance can be administered to an individual without causing undesirable biological effects or interacting in a harmful manner with any of the components of the composition in which it is contained.

[0192] As used herein, the term “pharmaceutical composition” refers to a mixture of at least one compound of the present invention with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspenders, thickeners, and / or excipients. Pharmaceutical compositions facilitate the administration of compounds to living organisms. Multiple techniques exist in the art for administering compounds, including but not limited to intravenous, oral, aerosol, parenteral, intraocular, pulmonary, and topical administration.

[0193] When used herein, “disease” or “disorder” refers to a condition that requires and / or is desired to be treated.

[0194] As used herein, “to treat,” “to treat,” or “treatment” means to improve a disease or disorder, for example, to slow, stop or reduce the progression of the disease or disorder or to reduce at least one of its clinical symptoms. For the purposes of this disclosure, improving a disease or disorder may include, but is not limited to, obtaining any beneficial or desired clinical outcome, including: reducing one or more symptoms; reducing the severity of the disease; preventing or delaying the spread of the disease (e.g., metastasis, e.g., to the lungs or lymph nodes); preventing or delaying the recurrence of the disease; slowing or delaying the progression of the disease; improving the disease state; inhibiting the disease or the progression of the disease; inhibiting or delaying the disease or its progression; stopping its progression; and remission (whether partial or whole).

[0195] The terms “individual” and “subject” are used interchangeably herein to refer to animals, e.g., mammals. The term “patient” includes humans and veterinary subjects. In some embodiments, methods are provided for treating mammals, including but not limited to humans, rodents, monkeys, cats, dogs, horses, cattle, pigs, sheep, goats, mammalian laboratory animals, mammalian farm animals, mammalian sports animals, and mammalian pets. Subjects may be male or female and may be of any preferred age, including infants, young, adolescent, adult, and elderly subjects. In some examples, “individual” or “subject” refers to an individual or subject requiring treatment for a disease or disorder. In some embodiments, the subject receiving treatment may be a patient, specifying the fact that the subject has an injury related to the treatment or is identified as being at sufficient risk of developing the disorder. In certain embodiments, the subject is a human, e.g., a human patient.

[0196] II. Targeted lipid particles (e.g., lentiviral vectors) This specification provides targeted lipid particles comprising a henipavirus F protein molecule or a biologically active moiety thereof, and a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or the biologically active moiety, and each of (i) and (ii) is exposed on the outer surface of the targeted lipid particle. In some embodiments, the binding domain is a single-domain antibody. In some embodiments, the binding domain is a single-stranded variable fragment. In certain embodiments, the provided lipid particles exhibit fusion activity mediated by the targeted envelope protein, which facilitates binding to target cells and facilitates the integration or fusion of the two lumens of the lipid particle and the target cell membrane, as well as the G protein or its biologically active moiety thereof, and the F glycoprotein.

[0197] This specification provides targeted lipid particles comprising a henipavirus F protein molecule or a biologically active moiety thereof, and a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof and (ii) a single-domain antibody (sdAb) variable domain, wherein the single-domain antibody is bound to the C-terminus of the G protein or the biologically active moiety, and each of (i) and (ii) is exposed on the outer surface of the targeted lipid particle. In certain embodiments, the provided lipid particles exhibit fusion activity mediated by the targeted envelope protein, which facilitates binding to target cells and facilitates the integration or fusion of the two lumens of the lipid particle and the target cell membrane, as well as the G protein or its biologically active moiety thereof, and the F glycoprotein.

[0198] In some of the various embodiments, the targeted lipid particles are viral particles or virus-like particles. In some embodiments, such targeted lipid particles contain viral nucleic acids, e.g., retroviral nucleic acids, e.g., lentiviral nucleic acids. In certain embodiments, any provided targeted lipid particles, e.g., viral particles or virus-like particles, are replication-deficient. In some embodiments, the targeted lipid particles are lentiviral vectors, and the lentiviral vectors are pseudotyped with henipavirus F protein and targeted envelope protein.

[0199] For example, this specification provides a pseudotyped lentiviral vector comprising a henipavirus F protein molecule or a biologically active moiety thereof, and a targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or a biologically active moiety thereof and (ii) a binding domain, wherein the binding domain is bound to the C-terminus of the G protein or the biologically active moiety, and each of (i) and (ii) is exposed on the outer surface of a targeted lipid particle. In some embodiments, the binding domain is a single-domain antibody. In some embodiments, the binding domain is a single-stranded variable fragment.

[0200] In some embodiments, the targeted lipid particles (e.g., targeted lentiviral vectors) provided herein have increased or greater targeted envelope protein expression compared to reference lipid particles (e.g., reference lentiviral vectors) that incorporate a similar envelope protein but are fused to an alternative targeting site other than the sdAb variable domain, such as a single-stranded variable fragment (scFv). In some embodiments, such targeted lipid particles are generated by pseudotyping of the lipid particles (e.g., lentiviral particles) after cotransfection of cells with transfer, envelope, and gag-pol plasmids.

[0201] In some embodiments, expression increases by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 40%, 500% or more compared to a reference lipid particle (e.g., a reference lentiviral vector), e.g., a reference lipid particle containing a similar envelope protein but fused to an scFv. In some examples, expression increases by 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold or more compared to a reference lipid particle (e.g., a reference lentiviral vector), e.g., a reference lipid particle containing a similar envelope protein but fused to an scFv. In some embodiments, expression can be assayed in vitro using flow cytometry, e.g., FAC. In some embodiments, expression may be expressed as the number or density of targeted envelope proteins on the surface of targeted lipid particles (e.g., targeted lentiviral vectors). In some embodiments, expression may be expressed as the mean fluorescence intensity (MFI) of surface expression of targeted envelope proteins on the surface of targeted lipid particles (e.g., targeted lentiviral vectors). In some embodiments, expression may be expressed as the percentage of lipid particles (e.g., lentiviral vectors) in a population that is surface-positive for targeted envelope proteins.

[0202] In some embodiments, in a population of targeted lipid particles (e.g., targeted lentiviral vectors), 50% or more than 50% of the lipid particles are surface-positive for the targeted envelope protein. For example, in a population of provided targeted lipid particles (e.g., targeted lentiviral vectors), 55% or more than 55%, 60% or more than 60%, 65% or more than 65%, 70% or more than 70%, and 75% or more than 75% of the cells in the population are surface-positive for the targeted envelope protein.

[0203] In some embodiments, the titer of targeted lipid particles after introduction into target cells, such as by transduction (e.g., transduction cells), increases compared to the titer of reference lipid particles (e.g., reference lentiviral vectors) to the same target cells, which incorporate similar envelope proteins but are fused to alternative targeting sites other than the sdAb variable domain, such as single-stranded variable fragments (scFvs). Typically, the alternative targeting sites recognize or bind to the same target molecules as the sdAb variable domain of the targeted envelope protein of the targeted lipid particles. In some embodiments, the titer increases by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 200%, 300%, 400%, 500% or more compared to the titer of reference lipid particles (e.g., reference lentiviral vectors), such as reference lipid particles containing similar envelope proteins but fused to scFvs. In some examples, the titer increases by 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 times or more compared to the titer of a reference lipid particle (e.g., a reference lentiviral vector), e.g., a reference lipid particle containing a similar envelope protein but fused to an scFv. In some embodiments, the titer of the targeted lipid particle in target cells (e.g., transduced cells) is 1 × 10⁻¹⁶ 6 Transduction units (TU) / mL or approximately 1 x 10⁻¹⁰ 6 The transduction unit (TU) / mL is greater than 2 × 10⁻¹⁰. For example, the titer of targeted lipid particles in target cells (e.g., transduction cells) is 2 × 10⁻¹⁰. 6 TU / mL or approximately 2 × 10 6 TU / mL or higher, 3 x 10 6 TU / mL or approximately 3 × 10 6 TU / mL or higher, 4 x 10 6 TU / mL or approximately 4 × 10 6 TU / mL or higher, 5 x 10 6 TU / mL or approximately 5 × 10 6 TU / mL or higher, 6 x 10 6 TU / mL or approximately 6 × 10 6 TU / mL or higher, 7 x 10 6TU / mL or approximately 7 × 10 6 TU / mL or higher, 8 x 10 6 TU / mL or approximately 8 x 10 6 TU / mL or higher, 9 x 10 6 TU / mL or approximately 9 x 10 6 Greater than TU / mL, or 1 × 10⁻⁶ 7 TU / mL or approximately 1 × 10⁻⁶ 7 It is greater than TU / mL.

[0204] A. Targeted envelope proteins (e.g., henipavirus-binding domain) In some embodiments, the targeted lipid particles (e.g., lentiviral vectors) include targeted envelope proteins exposed on the surface of the targeted lipid particles (e.g., lentiviral vectors).

[0205] In some embodiments, the targeted envelope protein contains a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and a binding domain that binds to a cell surface molecule on the target cell. In some embodiments, the binding domain is a single-domain antibody (sdAb). In some embodiments, the binding domain is a single-strand variable fragment (scFv). The binding domain may be linked directly or indirectly to the G protein. In certain embodiments, the binding domain is linked to the C-terminus (C-terminal amino acid) of the G protein or its biologically active moiety. Linking may be via a peptide linker, such as a flexible peptide linker.

[0206] 1. G protein In some embodiments, the targeted envelope protein comprises a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and a single-domain antibody (sdAb) variable domain or its biologically active moiety. In some embodiments, the sdAb binds to a cell surface molecule on the target cell. The sdAb variable domain may be linked directly or indirectly to the G protein. In certain embodiments, the sdAb variable domain is linked to the C-terminus (C-terminal amino acid) of the G protein or its biologically active moiety. Linking may be via a peptide linker, such as a flexible peptide linker.

[0207] In some embodiments, the binding domain (e.g., sdAb) binds to a cell surface antigen of a cell. In some embodiments, the cell surface antigen is characteristic of one type of cell. In some embodiments, the cell surface antigen is characteristic of multiple types of cells.

[0208] In some embodiments, the binding domain (e.g., sdAb) variable domain binds to a cell surface molecule or antigen. In some embodiments, the cell surface molecule is ASGR1, ASGR2, TM4SF5, CD8, CD4, or low-density lipoprotein receptor (LDL-R). In some embodiments, the cell surface molecule is ASGR1. In some embodiments, the cell surface molecule is ASGR2. In some embodiments, the cell surface molecule is TM4SF5. In some embodiments, the cell surface molecule is CD8. In some embodiments, the cell surface molecule is CD4. In some embodiments, the cell surface molecule is LDL-R.

[0209] In some embodiments, the G protein is the henipavirus G protein or its biologically active portion. In some embodiments, the henipavirus G protein is the hendra (HeV) virus G protein, the nipah (NiV) virus G protein (NiV-G), the cedar (CedPV) virus G protein, the Mojiang virus G protein, the bat paramyxovirus G protein or its biologically active portion. Table 3 provides non-limiting examples of G proteins.

[0210] The binding G protein is a type II transmembrane glycoprotein containing an N-terminal cytoplasmic tail (e.g., corresponding to amino acids 1-49 of SEQ ID NO: 9), a transmembrane domain (e.g., corresponding to amino acids 50-70 of SEQ ID NO: 9), an extracellular domain containing an extracellular stalk (e.g., corresponding to amino acids 71-187 of SEQ ID NO: 9), and a spherical head (e.g., corresponding to amino acids 188-602 of SEQ ID NO: 9). The N-terminal cytoplasmic domain is located within the inner lumen of the lipid bilayer, while the C-terminal portion is an extracellular domain exposed outside the lipid bilayer. The stalk region in the C-terminal region (e.g., corresponding to amino acids 159-167 of NiV-G) has been shown to be involved in interaction with the F protein and induction of F protein fusion (Liu et al. 2015 J of Virology 89:1838). In wild-type G proteins, the globular head mediates receptor binding to henipavirus entry receptors ephrin B2 and ephrin B3 but is unnecessary for membrane fusion (Brandel-Tretheway et al. Journal of Virology. 2019. 93(13)e00577-19). In certain embodiments herein, the directivity of a G protein is modified by ligation of the G protein or a biologically active fragment (e.g., a cytoplasmic cleavage) to the sdAb variable domain. Binding of the G protein to a binding partner can induce fusion mediated by a compatible F protein or its biologically active moiety. The G protein sequences disclosed herein are primarily disclosed as expressed sequences containing the N-terminal methionine required for translation initiation. Since such N-terminal methionine is generally cleaved during or after translation, the mature protein sequences for all G protein sequences disclosed herein are also intended to lack the N-terminal methionine.

[0211] G glycoproteins are highly conserved among henipavirus species. For example, the G proteins of NiV and HeV viruses share 79% amino acid identity. Studies have shown high compatibility between G proteins and F proteins of different species, as demonstrated by heteromorphic fusion activation (Brandel-Tretheway et al. Journal of Virology. 2019). As further described below, retargeted lipid particles may contain heteromorphic G and F proteins from different species.

[0212] (Table 3) Henipavirus protein G sequence clusters Column 1: The Genbank ID contains the Genbank ID for the entire viral genome sequence, which is the centroid sequence of the cluster. Column 2: The nucleotides in CDS provide the nucleotides corresponding to the CDS of genes throughout the genome. Column 3: Provides the complete gene name, including the Genbank ID, virus species, strain, and protein name. Column 4: A sequence, which provides the amino acid sequence of a gene. Column 5: Number of arrays / cluster, which provides the number of centroid arrays and the number of arrays to cluster. Column 6: Provides the array index for the sequence described. TIFF0007887223000003.tif156165TIFF0007887223000004.tif205165TIFF0007887223000005.tif196165TIFF0007887223000006.tif162165

[0213] In some embodiments, the G protein has a sequence shown in any of SEQ ID NOs: 9, 18, 28, 29, 30, 31, 44, 52, or 54-56, or a sequence with at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 8 The functionally active variant or biologically active moiety has a sequence that is identical to 7%, at least 88% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, at least 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%. In certain embodiments, the G protein or functionally active variant or biologically active moiety is a protein that maintains fusion activity in combination with a henipavirus F protein, e.g., the F protein shown in section IB (e.g., NiV-F or HeV-F). Fusion activity includes the activity of the G protein in combination with the henipavirus F protein to promote or facilitate the fusion of two membrane lumens, for example, the lumen of a targeted lipid particle having henipavirus F and G proteins embedded in its lipid bilayer, and the cytoplasm of a target cell, for example, a cell containing a surface receptor or molecule recognized or bound by a targeted envelope protein. In some embodiments, the F protein and G protein are from the same henipavirus species (e.g., NiV-G and NiV-F). In some embodiments, the F protein and G protein are from different henipavirus species (e.g., NiV-G and HeV-F).

[0214] In certain embodiments, the G protein has the amino acid sequence shown in SEQ ID NO: 9, SEQ ID NO: 28, SEQ ID NO: 18, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 44, SEQ ID NO: 52, or SEQ ID NO: 54-56, or is a functionally active variant thereof or a biologically active portion thereof that retains fusion activity. In some embodiments, the functionally active variant comprises an amino acid sequence having at least 80% or about 80%, at least 85% or about 85%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with the henipavirus F protein (e.g., NiV-F or HeV-F), and retains fusion activity in combination with the henipavirus F protein (e.g., NiV-F or HeV-F). In some embodiments, the biologically active portion has an amino acid sequence having at least 80% or about 80%, at least 85% or about 85%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 9, SEQ ID NO: 28, SEQ ID NO: 18, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 44, SEQ ID NO: 52, or SEQ ID NO: 54-56, and retains fusion activity in combination with henipavirus F protein (e.g., NiV-F or HeV-F).

[0215] References to maintaining fusion activity include levels or degrees of binding of the corresponding wild-type G protein between 10% or about 10% and 150% or about 150%, or more, as shown in SEQ ID NOs. 9, SEQ ID NOs. 28, SEQ ID NOs. 18, SEQ ID NOs. 30, SEQ ID NOs. 31, SEQ ID NOs. 44, SEQ ID NOs. 52 or SEQ ID NOs. 54-56, for example, at least or at least about 10% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or at least about 15% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or at least about 20% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or at least about 25% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or at least about 30% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or at least about 35% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or at least about 40% of the level or degree of fusion activity of the corresponding wild-type G protein. For example, at least or about 45% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 50% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 55% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 60% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 65% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 70% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 75% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 80% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 85% of the level or degree of fusion activity of the corresponding wild-type G protein, for example, at least or about 90% of the level or degree of fusion activity of the corresponding wild-type G protein,For example, the activity (combined with Nipah virus F protein) includes at least or about 95% of the level or degree of fusion activity of the corresponding wild-type G protein, at least or about 100%, or at least about 120% of the level or degree of fusion activity of the corresponding wild-type G protein.

[0216] In some embodiments, the G protein is a mutant G protein which is a functionally active variant or biologically active moiety containing one or more amino acid mutations, e.g., one or more amino acid insertions, deletions, substitutions, or cleavages. In some embodiments, the mutations described herein relate to amino acid insertions, deletions, substitutions, or cleavages of amino acids compared to a reference G protein sequence. In some embodiments, the reference G protein sequence is the wild-type sequence of the G protein or its biologically active moiety. In some embodiments, the functionally active variant or biologically active moiety is a variant of the wild-type Hendra (HeV) virus G protein, wild-type Nipah (NiV) virus G protein (NiV-G), wild-type Cedar (CedPV) virus G protein, wild-type Mojiang virus G protein, wild-type bat paramyxovirus G protein or its biologically active moiety. In some embodiments, the wild-type G protein has the sequence shown in any one of SEQ ID NOs: 9, 18, 28, 29, 30, 31, SEQ ID NOs: 44, SEQ ID NOs: 52, or SEQ ID NOs: 54-56.

[0217] In some embodiments, the G protein is a mutant G protein, which is a biologically active portion that is a fragment cleaved at the N-terminus and / or C-terminus of a wild-type Hendra (HeV) virus G protein, wild-type Nipah (NiV) virus G protein (NiV-G), wild-type Cedar (CedPV) virus G protein, wild-type Mojiang virus G protein, or wild-type bat paramyxovirus G protein. In certain embodiments, the cleavage is an N-terminal cleavage of all or part of the cytoplasmic domain. In some embodiments, the mutant G protein is a biologically active portion that is cleaved and lacks up to 49 consecutive amino acid residues at or near the N-terminus of a wild-type G protein, such as the wild-type G protein shown in SEQ ID NOs. 9, 18, 28, 29, 30, 31, SEQ ID NOs. 44, SEQ ID NOs. 52, or SEQ ID NOs. 54-56. In some embodiments, the mutant F protein is cleaved and lacks up to 49 consecutive amino acids at the N-terminus of the wild-type G protein, for example, up to 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 30, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 consecutive amino acid.

[0218] In some embodiments, the G protein is the wild-type Nipah virus G (NiV-G) protein or the Hendra virus G protein, or a functionally active variant or biologically active portion thereof. In some embodiments, the G protein is the NiV-G protein having the sequence shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, or at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, at least 8 It is a functional variant or biologically active moiety having an amino acid sequence having sequence identity of 8% or about 88%, at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, and at least 99% or about 99%.

[0219] In some embodiments, the G protein is a mutant NiV-G protein, which is the biologically active portion of wild-type NiV-G. In some embodiments, the biologically active portion is a fragment cleaved at the N-terminus. In some embodiments, the mutant NiV-G protein is cleaved and contains up to 5 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 6 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 7 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 8 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 9 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), and up to 10 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44). 0.0 amino acid residues, up to 11 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO 9, SEQ ID NO 28, or SEQ ID NO 44), up to 12 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO 9, SEQ ID NO 28, or SEQ ID NO 44), up to 13 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO 9, SEQ ID NO 28, or SEQ ID NO 44), up to 14 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO 9, SEQ ID NO 28, or SEQ ID NO 44), up to 15 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO 9, SEQ ID NO 28, or SEQ ID NO 44), up to 16 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO 9, SEQ ID NO 28, or SEQ ID NO 44), wild-type NiV-G protein (SEQ ID NO 9,Up to 17 consecutive amino acid residues at or near the N-terminus of SEQ ID NO: 28 or SEQ ID NO: 44, up to 18 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44), up to 19 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44), up to 20 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44) The base, up to 21 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 22 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 23 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) or up to 24 consecutive amino acid residues near or at the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 25 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 26 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 27 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 44) Up to 28 consecutive amino acid residues at or near the N-terminus of (SEQ ID NO. 28 or SEQ ID NO. 44), up to 29 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO. 9, SEQ ID NO. 28 or SEQ ID NO. 44), up to 30 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO. 9, SEQ ID NO. 28 or SEQ ID NO. 44), up to 31 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO. 9, SEQ ID NO. 28 or SEQ ID NO. 44),Up to 32 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 33 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 34 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) Up to 35 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 36 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 37 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 38 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 39 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 40 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 41 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to It lacks 42 consecutive amino acid residues, up to 43 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), up to 44 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), or up to 45 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44).

[0220] In some embodiments, the NiV-G protein is a biologically active portion that does not contain a cytoplasmic domain. In some embodiments, the NiV-G protein without a cytoplasmic domain is encoded by SEQ ID NO: 32.

[0221] In some embodiments, the mutant NiV-G protein contains the sequence shown in any of SEQ ID NOs: 10-15, 35-40, 45-50, 22, 53 or SEQ ID NO: 32, or at least 80% or 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least It is a functional variant having an amino acid sequence with sequence identity of 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0222] In some embodiments, the mutant NiV-G protein is cleaved at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), for example, as shown in SEQ ID NO: 10 or SEQ ID NO: 10, with at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. , a functional variant having sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in Sequence ID No. 35. or sequence number 35 and at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%, at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93% , a functional variant having sequence identity of at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, the one shown in SEQ ID NO: 45 or SEQ ID NO: 45 with at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%,It has a functional variant having sequence identity of at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%. In some embodiments, the mutant NiV-G protein is cleaved by at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, and at least 94%. or a functional variant having sequence identity of approximately 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%, or, for example, as shown in sequence number 36 or sequence number 36 with at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%,A functional variant having sequence identity of at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in SEQ ID NO: 46 or at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%, It has a functional variant with sequence identity of at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0223] In some embodiments, the mutant NiV-G protein is cleaved at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), for example, as shown in SEQ ID NO: 12 or SEQ ID NO: 12, by at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, A functional variant having an amino acid sequence with sequence identity of at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, as shown in, for example, SEQ ID NO: 37 Those that are included or number 37 and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 9 Its functional variant having sequence identity of 3%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99%, or, for example, as shown in SEQ ID NO: 47 or at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%,It has a functional variant with sequence identity of at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%. In some embodiments, the mutant NiV-G protein is cleaved at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), for example, as shown in SEQ ID NO: 13 or SEQ ID NO: 13, and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. A functional variant having sequence identity of %, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in SEQ ID NO: 38. or sequence number 38 and at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%, at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93% , a functional variant having sequence identity of at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in sequence number 48 or sequence number 48 with at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%,It has a functional variant with sequence identity of at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%. In some embodiments, the mutant NiV-G protein is cleaved at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), for example, as shown in SEQ ID NO: 14 or SEQ ID NO: 14, and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. A functional variant having sequence identity of %, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in sequence number 39. or sequence number 39 and at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%, at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93% , a functional variant having sequence identity of at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in sequence number 49 or sequence number 49 with at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%,It has a functional variant with sequence identity of at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%. In some embodiments, the mutant NiV-G protein is cleaved at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44) by at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%. A functional variant having sequence identity of %, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in sequence number 40. or sequence number 40 and at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%, at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, or at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93% , a functional variant having sequence identity of at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%, or, for example, as shown in SEQ ID NO: 50 or at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%,It has a functional variant with sequence identity of at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or approximately 88%, at least 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%. In some embodiments, the mutant NiV-G protein is cleaved at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), for example, as shown in SEQ ID NO: 22 or SEQ ID NO: 22, and at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98% , or a functional variant thereof having at least 99% or approximately 99% sequence identity, or, for example, the one shown in SEQ ID NO: 53 or SEQ ID NO: 53 with at least 80% or approximately 80%, at least 81% or approximately 81%, at least 82% or approximately 82%, at least 83% or approximately 83%, 84% or approximately 84%, at least 85% or approximately 85%, at least 86% or approximately 86%, or at least 87% or approximately 87%, at least 88% or It has a functional variant having sequence identity of approximately 88%, or at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%. In some embodiments, the mutant HiV-G protein is at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90% , lacking the N-terminal cytoplasmic domain of the wild-type HiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44), as indicated by its functional variant having at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99% sequence identity.

[0224] In some embodiments, the mutant G protein is a mutant HeV-G protein having the sequence shown in SEQ ID NO: 18 or 52, or at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, at least 84% or about 84%, 85% or about 85%, at least 86% or about 86%, at least 87% or about 87%, 88% or about 88%, at least It is a functional variant or biologically active moiety having an amino acid sequence having sequence identity of 89% or approximately 89%, at least 90% or approximately 90%, at least 91% or approximately 91%, at least 92% or approximately 92%, at least 93% or approximately 93%, at least 94% or approximately 94%, at least 95% or approximately 95%, 96% or approximately 96%, at least 97% or approximately 97%, at least 98% or approximately 98%, or at least 99% or approximately 99%.

[0225] In some embodiments, the G protein is a mutant HeV-G protein, which is the biologically active portion of wild-type HeV-G. In some embodiments, the biologically active portion is a fragment cleaved at the N-terminus. In some embodiments, the mutant HeV-G protein is cleaved and has up to 5 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 6 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 7 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 8 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 9 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 10 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 11 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein ( Up to 12 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 13 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 14 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 15 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 16 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 17 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 18 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 19 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52),Up to 20 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 21 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 22 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 23 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52) Up to 24 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 25 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 26 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 27 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), and the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52) Up to 28 consecutive amino acid residues at or near the end, up to 29 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 30 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 31 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), and at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52). Up to 32 consecutive amino acid residues, up to 33 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 34 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 35 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 36 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52),Up to 37 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 38 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 39 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 40 consecutive amino acid residues at or near the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52), at the N-terminus of wild-type HeV-G protein (SEQ ID NO: 18 or 52) It lacks up to 41 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 42 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 43 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), up to 44 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52), or up to 45 consecutive amino acid residues at or near the N-terminus of the wild-type HeV-G protein (SEQ ID NO: 18 or 52). In some embodiments, the HeV-G protein is a biologically active portion that does not contain a cytoplasmic domain. In some embodiments, the mutant HeV-G protein has a functional variant that has at least 80% or about 80%, at least 81% or about 81%, at least 82% or about 82%, at least 83% or about 83%, 84% or about 84%, at least 85% or about 85%, at least 86% or about 86%, or at least 87% or about 87%, at least 88% or about 88%, or at least 89% or about 89%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 33 or SEQ ID NO: 33, as shown in its functional variant.It lacks the N-terminal cytoplasmic domain of the wild-type HeV-G protein (SEQ ID NO: 18 or 52).

[0226] In some embodiments, the G protein or a functionally active variant or biologically active moiety thereof binds to ephrin B2 or ephrin B3. In some embodiments, the G protein is a functionally active variant or biologically active moiety thereof having the amino acid sequence shown in any one of SEQ ID NOs: 9, SEQ ID NOs: 18 or 28, SEQ ID NOs: 29, SEQ ID NOs: 44, SEQ ID NOs: 30 or 31, or capable of binding to ephrin B2 or ephrin B3. In some embodiments, the functionally active variant or biologically active moiety has an amino acid sequence having at least about 80%, at least about 85%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31, or a functionally active variant or biologically active moiety thereof, and retains binding to ephrin B2 or B3. References to retain binding to ephrin B2 or B3 include at least or about 5% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 10% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 15% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9,20% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 25% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 30% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 35% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, 40% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 45% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 50% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 55% of the binding level or degree of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44,60% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 30 or SEQ ID NO: 31; 65% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; 70% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31; for example, at least or at least about 75% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 1 At least or about 80% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 8 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31, for example, at least or about 85% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31, for example, at least or about 90% of the level or degree of binding of the corresponding wild-type G protein, or its functionally active variant or biologically active moiety, as shown in SEQ ID NO: 9, SEQ ID NO: 18 or SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 44, SEQ ID NO: 30 or SEQ ID NO: 31,or the binding includes at least or about 95% of the level or degree of binding of the functionally active variant or biologically active moiety thereof. In some embodiments, the G protein is NiV-G or a functionally active variant or biologically active moiety thereof that binds to ephrin B2 or ephrin B3. In some embodiments, NiV-G is a functionally active variant or biologically active moiety thereof having the amino acid sequence shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, or that can bind to ephrin B2 or ephrin B3. In some embodiments, a functionally active variant or biologically active moiety has an amino acid sequence having at least about 80%, at least about 85%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NOs. Exemplary biologically active moieties include, for example, N-terminal cleavage variants lacking all or part of the cytoplasmic domain, e.g., one or more, e.g., 1 to 49 consecutive N-terminal amino acid residues, as shown in any one of SEQ ID NOs. 10-15, 35-40, 45-50, and 32. References to retaining binding to ephrin B2 or B3 include at least or about 5% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 10% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 15% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 20% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44,25% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 28 or SEQ ID NO: 44, 30% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 35% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 40% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 45% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 50% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 55% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44, 60% of the level or degree of binding of the corresponding wild-type NiV-G as shown in SEQ ID NO: 9, SEQ ID NO: 28 or SEQ ID NO: 44 The binding includes levels or degrees of 65% of the corresponding wild-type NiV-G binding, 70% of the corresponding wild-type NiV-G binding level or degree as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, for example, at least or at least about 75% of the corresponding wild-type NiV-G binding level or degree as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, for example, at least or at least about 80% of the corresponding wild-type NiV-G binding level or degree as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, for example, at least or at least about 85% of the corresponding wild-type NiV-G binding level or degree as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, for example, at least or at least about 90% of the corresponding wild-type NiV-G binding level or degree as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, or for example, at least or at least about 95% of the corresponding wild-type NiV-G binding level or degree as shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44.

[0227] In some embodiments, the G protein is HeV-G or a functionally active variant or bioactive moiety thereof that binds to ephrin B2 or ephrin B3. In some embodiments, HeV-G is a functionally active variant or bioactive moiety thereof having the amino acid sequence shown in SEQ ID NO: 18 or 52, or capable of binding to ephrin B2 or ephrin B3. In some embodiments, a functionally active variant or biologically active moiety has an amino acid sequence having at least about 80%, at least about 85%, at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% sequence identity with SEQ ID NO: 18 or 52, and retains binding to ephrin B2 or B3. Exemplary biologically active moieties include, for example, N-terminal cleavage variants lacking all or part of the cytoplasmic domain, e.g., one or more, e.g., 1 to 49 consecutive N-terminal amino acid residues, as shown in any one of SEQ ID NOs: 33. References to retain binding to ephrin B2 or B3 include at least or at least about 5% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 10% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 15% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 20% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 25% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 30% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 35% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52,40% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 45% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 50% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 55% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 60% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 65% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, 70% of the binding level or degree of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, for example. The binding includes at least or about 75% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, for example, at least or about 80% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, for example, at least or about 85% of the level or degree of binding of the corresponding wild-type NIV-G as shown in SEQ ID NO: 18 or 52, for example, at least or about 90% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52, or for example, at least or about 95% of the level or degree of binding of the corresponding wild-type HeV-G as shown in SEQ ID NO: 18 or 52.

[0228] In some embodiments, the G protein or its biologically active moiety is a mutant G protein that exhibits reduced binding to the native binding partner of the wild-type G protein. In some embodiments, the mutant G protein or its biologically active moiety is a mutant of wild-type Niv-G and exhibits reduced binding to one or both of the native binding partners ephrin B2 or ephrin B3. In some embodiments, the mutant G protein or its biologically active moiety, for example, the mutant NiV-G protein, exhibits reduced binding to its native binding partner. In some embodiments, the reduced binding to ephrin B2 or ephrin B3 is reduced by more than 5% or about 5%, 10% or about 10%, 15% or about 15%, 20% or about 20%, 25% or about 25%, 30% or about 30%, 40% or about 40%, 50% or about 50%, 60% or about 60%, 70% or about 70%, 80% or about 80%, 90% or about 90%, or 100% or about 100%.

[0229] In some embodiments, the mutations described herein can improve transduction efficiency. In some embodiments, the mutations described herein enable specific targeting of other desired cell types other than ephrin B2 or ephrin B3. In some embodiments, the mutations described herein at least partially prevent binding to at least one intrinsic receptor, for example, by reducing binding to at least one of ephrin B2 or ephrin B3. In some embodiments, the mutations described herein interfere with intrinsic receptor recognition.

[0230] In some embodiments, the G protein contains one or more amino acid substitutions in residues involved in interactions with one or both of ephrin B2 and ephrin B3. In some embodiments, the amino acid substitutions correspond to mutations E501A, W504A, Q530A, and E533A, with reference to the numbering shown in Sequence ID No. 28.

[0231] In some embodiments, the G protein is a mutant G protein containing one or more amino acid substitutions selected from the group consisting of E501A, W504A, Q530A, and E533A, with reference to the numbering shown in SEQ ID NO: 28, and its biologically active portion contains an N-terminal cleavage. In some embodiments, the mutant NiV-G protein or its biologically active portion is cleaved, with up to 5 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 6 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 7 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 8 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 9 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), up to 10 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), and at the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28) 11 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 12 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 13 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 14 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), up to 15 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 16 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 17 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), 18 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28),19 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), up to 20 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 21 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 22 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 23 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28) 24 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), up to 25 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 26 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 27 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), at the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28) 28 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 29 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), up to 30 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), up to 31 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 32 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 33 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 34 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), 35 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), up to 36 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28), up to 37 consecutive amino acid residues at or near the N-terminus of wild-type NiV-G protein (SEQ ID NO: 28),It lacks up to 38 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), up to 39 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28), or up to 40 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 28).

[0232] In some embodiments, the mutant NiV-G protein has an amino acid sequence that is at least 90% or about 90%, at least 91% or about 91%, at least 92% or about 92%, at least 93% or about 93%, at least 94% or about 94%, at least 95% or about 95%, 96% or about 96%, at least 97% or about 97%, at least 98% or about 98%, or at least 99% or about 99% of sequence identity with SEQ ID NO: 16 or 51. In certain embodiments, the G protein has an amino acid sequence that is at least 99% or about 99% of the sequence shown in SEQ ID NO: 16 or 51.

[0233] In some embodiments, the targeted envelope protein contains a G protein or a functionally active variant or a biologically active moiety and an sdAb variable domain, and the targeted envelope protein exhibits increased binding to a molecule different from the native binding partner of the wild-type G protein. In some embodiments, the molecule may be a protein expressed on the surface of a desired target cell. In some embodiments, the increased binding to the other molecule increases by 25% or about 25%, 30% or about 30%, 40% or about 40%, 50% or about 50%, 60% or about 60%, 70% or about 70%, 80% or about 80%, 90% or about 90%, or 100% or about 100%. In certain embodiments, the binding confers a retargeted binding compared to the binding of the wild-type G protein, in which case a new or different binding activity is conferred.

[0234] 2. Joint Domain In some embodiments, the binding domain may be any substance that binds to a cell surface molecule on a target cell. In some embodiments, the binding domain may be an antibody or an antibody moiety or fragment.

[0235] The binding domain can be modified to have different binding strengths. For example, scFv and antibodies with varying binding strengths can be used to modify the fusion activity of chimeric binding proteins to cells showing large or small amounts of the target antigen. For example, DARPin with different affinities can be used to modify the fusion activity to cells showing large or small amounts of the target antigen. The binding domain can also be modified to target different regions on the target ligand, which affects the rate of fusion with cells presenting the target.

[0236] The binding domains include: humanized antibody molecules, intact IgA, IgG, IgE, or IgM antibodies; bi- or multispecific antibodies (e.g., Zybodies®); antibody fragments (e.g., Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments, and isolated CDRs or sets thereof); single-stranded Fvs; polypeptide-Fc fusions; single-domain antibodies (e.g., shark single-domain antibodies, e.g., IgNAR or fragments thereof); camel-like antibodies; masked antibodies (e.g., Probodies®); small module immunotherapies ("SMIPs™"); single-stranded or tandem diaphragms. This may include Bodies (TandAb(registered trademark)); VHH; Anticalins(registered trademark); Nanobodies(registered trademark); Minibodies; BiTE(registered trademark); Ankyrin Repeat Protein or DARPIN(registered trademark); Avimer(registered trademark); DART; TCR-like antibodies; Adnectin(registered trademark); Affilin(registered trademark); Trans-bodies(registered trademark); Affibodies(registered trademark); TrimerX(registered trademark); MicroProteins; Fynomers(registered trademark), Centyrins(registered trademark); and KALBITOR(registered trademark). Targeting sites may also include antibodies or their antigen-binding fragments (e.g., Fab, Fab', F(ab')2, Fv fragment, scFv antibody fragment, disulfide-linked Fv(sdFv), Fd fragment consisting of VH and CH1 domains, linear antibodies, single-domain antibodies, e.g., sdAb(either VL or VH), nanobodies, or camelid VHH domains), antigen-binding fibronectin type III (Fn3) skeletons, e.g., fibronectin polypeptide minibodies, ligands, cytokines, chemokines, or T cell receptors (TCRs).

[0237] In some embodiments, the binding domain is a single-stranded molecule. In some embodiments, the binding domain is a single-domain antibody. In some embodiments, the binding domain is a single-stranded variable fragment. In certain embodiments, the binding domain contains a human or humanized antibody variable sequence(s).

[0238] In some embodiments, the binding domain is a single-domain antibody. In some embodiments, the single-domain antibody may be human or humanized. In some embodiments, the single-domain antibody or its portion is naturally occurring. In some embodiments, the single-domain antibody or its portion is synthetic.

[0239] In some embodiments, a single-domain antibody is an antibody in which the complementarity-determining region is part of a single-domain polypeptide. In some embodiments, a single-domain antibody is an antibody variable domain consisting only of a heavy chain. In some embodiments, a single-domain antibody does not contain a light chain.

[0240] In some embodiments, heavy-chain antibodies lacking a light chain are referred to as VHH. In some embodiments, single-domain antibodies have a molecular weight of 12-15 kDa. In some embodiments, single-domain antibodies include camelid antibodies or shark antibodies. In some embodiments, single-domain antibody molecules are derived from antibodies established in camelid species, such as camels, llamas, dromedary camels, alpacas, vicuñas, and guanacos. In some embodiments, single-domain antibodies are referred to as immunoglobulin neoantigen receptors (IgNARs) and are derived from cartilaginous fish. In some embodiments, single-domain antibodies are produced by splitting the dimeric variable domain of human or mouse IgG into monomers and camelidizing key residues.

[0241] In some embodiments, single-domain antibodies may be generated from a phage display library. In some embodiments, the phage display library is generated from a camelid VHH repertoire immunized with various antigens, as described in Arbabi et al., FEBS Letters, 414, 521-526 (1997), Lauwereys et al., EMBO J., 17, 3512-3520 (1998), and Decanniere et al., Structure, 7, 361-370 (1999). In some embodiments, the phage display library is generated by including antibody fragments from unimmunized camelids. In some embodiments, a single-domain antibody library of human single-domain antibodies is generated synthetically by introducing diversity into one or more scaffolds.

[0242] In some embodiments, the C-terminus of a single-domain antibody is bound to the C-terminus of a G protein or its biologically active portion. In some embodiments, the N-terminus of a single-domain antibody is exposed on the outer surface of a lipid bilayer. In some embodiments, the N-terminus of a single-domain antibody binds to cell surface molecules of a target cell. In some embodiments, the single-domain antibody specifically binds to cell surface molecules present on the target cell. In some embodiments, the cell surface molecules are proteins, glycans, lipids, or low molecular weight molecules.

[0243] In some embodiments, the cell surface molecule of the target cell is an antigen or a part thereof. In some embodiments, the single-domain antibody or a part thereof is an antibody having a single monomeric domain antigen-binding / recognition domain that can selectively bind to a specific antigen. In some embodiments, the single-domain antibody binds to an antigen present on the target cell.

[0244] Exemplary cells include polymorphonuclear cells (also known as PMNs, PMLs, PMNLs, or granulocytes), stem cells, embryonic ...

Claims

1. Targeted lipid particles, (a) A lipid bilayer surrounding the lumen, (b) Henipavirus F protein molecule or its biologically active portion, (c) A targeted envelope protein comprising (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active moiety and (ii) a single-domain antibody (sdAb) variable domain, wherein the sdAb variable domain is bound to the C-terminus of the G protein or its biologically active moiety, and / or the sdAb is bound to the G protein or its biologically active moiety via a peptide linker, and the sdAb binds to a cell surface molecule of a target cell. The targeted lipid particle comprises the F protein molecule or its biologically active portion and the targeted envelope protein, wherein the F protein molecule or its biologically active portion and the targeted envelope protein are embedded in a lipid bilayer.

2. The cell surface molecule is a protein, glycan, lipid or low molecular weight molecule; and / or The target cells are selected from the group consisting of tumor-infiltrating lymphocytes, T cells, neoplasms or tumor cells, virus-infected cells, stem cells, central nervous system (CNS) cells, hematopoietic stem cells (HSCs), liver cells, or fully differentiated cells. Targeted lipid particles according to claim 1.

3. The targeted lipid particles according to claim 1 or 2, wherein the target cells are selected from the group consisting of CD3+ T cells, CD4+ T cells, CD8+ T cells, hepatocytes, hematopoietic stem cells, CD34+ hematopoietic stem cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD105+ endothelial cells, B cells, CD20+ B cells, CD19+ B cells, cancer cells, CD133+ cancer cells, EpCAM+ cancer cells, CD19+ cancer cells, Her2 / Neu+ cancer cells, GluA2+ neurons, GluA4+ neurons, NKG2D+ natural killer cells, SLC1A3+ astrocytic cells, SLC7A10+ adipocytes, or CD30+ lung epithelial cells.

4. The single-domain antibody binds to an antigen or a portion thereof present on hepatocytes; and / or the cell surface molecule or antigen is selected from the group consisting of ASGR1, ASGR2, and TM4SF; or The cell surface molecule or antigen is CD8 or CD4; or The cell surface molecule or antigen is LDL-R. Targeted lipid particles according to any one of claims 1 to 3.

5. The lipid particles are lentiviral vectors; and / or the lipid bilayer is a viral envelope or contains one. Targeted lipid particles according to any one of claims 1 to 4.

6. The targeted lipid particle according to any one of claims 1 to 5, wherein the peptide linker has a length of up to 65 amino acids.

7. The peptide linker comprises a length of 2 to 65 amino acids; The targeted lipid particle according to claim 6, wherein the flexible linker comprises GS, GGS, GGGGS (SEQ ID NO: 43), GGGGGS (SEQ ID NO: 41), or a combination thereof, or the peptide linker comprises (GGS)n where n is 1 to 10, (GGGGGS)n where n is 1 to 10 (SEQ ID NO: 42), or (GGGGGS)n where n is 1 to 6 (SEQ ID NO: 27).

8. The targeted lipid particle according to any one of claims 1 to 7, wherein the G protein or its biologically active portion is wild-type Nipah virus G (NiV-G) protein or Hendra virus G protein, or its biologically active portion.

9. The NiV-G protein or its biologically active portion includes the amino acid sequence shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44, or an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44; The NiV-G protein is a cleaved and biologically active region lacking up to 40 consecutive amino acid residues at or near the N-terminus of the wild-type NiV-G protein (SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO: 44); and / or The NiV-G protein is a biologically active portion cleaved at the N-terminus of wild-type NiV-G, and has an amino acid sequence that is at least 90% identical to the sequence shown in SEQ ID NOs: 10-15, 35-40, or 45-50, or the sequence shown in SEQ ID NOs: 10-15, 35-40, or 45-50. Targeted lipid particles according to claim 8.

10. The targeted lipid particle according to claim 8 or 9, wherein the wild-type NiV-G protein is shown in SEQ ID NO: 9, SEQ ID NO: 28, or SEQ ID NO:

44.

11. The targeted lipid particle according to any one of claims 1 to 10, wherein the G protein or its biologically active portion is a mutant NiV-G protein exhibiting reduced binding to ephrin B2 or ephrin B3.

12. The mutant NiV-G protein or its biologically active portion One or more amino acid substitutions corresponding to amino acid substitutions selected from the group consisting of E501A, W504A, Q530A, and E533A, with reference to the numbering shown in Sequence ID No. 28; Refer to the numbering shown in Sequence ID No. 28 for amino acid substitutions E501A, W504A, Q530A and E533A; and / or The amino acid sequence shown in SEQ ID NO: 16, or an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 16, Targeted lipid particles according to claim 11, comprising:

13. The targeted lipid particle according to any one of claims 1 to 12, wherein the F protein or its biologically active portion is wild-type Nipah virus F (NiV-F) protein or Hendra virus F protein, or its biologically active portion.

14. (a) The NiV-F protein or its biologically active portion comprises the amino acid sequence shown in SEQ ID NO: 2, or an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 2; or (b) The NiV-F protein is i) A 20-amino acid cleavage at or near the C-terminus of the wild-type NiV-F protein (SEQ ID NO: 2), and / or ii) Point mutations in N-linked glycosylation sites It is the biologically active part that includes; or (c) The NiV-F protein is the biologically active portion of the wild-type NiV-F protein (SEQ ID NO: 2) having a 22-amino acid cleavage at or near its C-terminus. Targeted lipid particles according to claim 13.

15. The NiV-F protein or the biologically active portion has an amino acid sequence that is the sequence shown in SEQ ID NO: 5 or SEQ ID NO: 7, or has at least 90% sequence identity with SEQ ID NO: 5 or SEQ ID NO: 7; or The NiV-F protein or the biologically active portion has an amino acid sequence encoded by a nucleotide sequence encoding the sequence shown in SEQ ID NO: 8 or SEQ ID NO: 23, or a sequence having at least 90% sequence identity with SEQ ID NO: 8 or SEQ ID NO:

23. Targeted lipid particles according to claim 13 or 14.

16. The targeted lipid particle according to any one of claims 1 to 15, wherein the F protein comprises the sequence shown in SEQ ID NO: 23, and the G protein comprises the sequence shown in SEQ ID NO:

16.

17. The targeted lipid particle according to any one of claims 1 to 16, wherein the lipid particle further comprises an exogenous substance.

18. The exogenous substance is present in the lumen; The aforementioned exogenous substance is a protein or nucleic acid; The exogenous substance is a nucleic acid that codes for cargo for delivery to the target cell, and optionally the target cell is a T cell; The aforementioned exogenous substance codes for a therapeutic agent or a diagnostic agent; The exogenous substance encodes a membrane protein; and / or The aforementioned exogenous substance is a nucleic acid containing a payload gene for correcting a gene defect. Targeted lipid particles according to claim 17.

19. The targeted lipid particle according to claim 18, wherein the nucleic acid is DNA or RNA.

20. The targeted lipid particle according to claim 18 or 19, wherein the membrane protein is an antigen receptor for targeting cells expressed by or associated with a disease or pathological condition.

21. The targeted lipid particle according to any one of claims 18 to 20, wherein the membrane protein is a chimeric antigen receptor (CAR).

22. The targeted lipid particle according to any one of claims 18 to 21, wherein the gene deficiency is a gene deficiency in the target cell and / or the gene deficiency is related to liver cells or hepatocytes.

23. (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and (ii) a single-domain antibody (sdAb) variable domain that binds to a cell surface molecule present on a target cell, wherein the sdAb variable domain is bound to the C-terminus of the G protein or its biologically active portion, Polynucleotides containing these nucleotides.

24. (i) a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and (ii) a single-domain antibody (sdAb) variable domain that binds to a cell surface molecule present on a target cell, wherein the sdAb variable domain is bound to the C-terminus of the G protein or its biologically active portion; and Henipavirus F protein molecule or nucleic acid sequence encoding its biologically active portion, Polynucleotides containing these nucleotides.

25. A vector or plasmid comprising the polynucleotide described in claim 23.

26. A vector or plasmid comprising the polynucleotide described in Claim 24.

27. A cell comprising the polynucleotide described in claim 23, or the vector or plasmid described in claim 25.

28. A cell comprising the polynucleotide according to claim 24, or the vector or plasmid according to claim 26.

29. The cell according to claim 27, wherein the cell is a producer cell.

30. The cell according to claim 28, wherein the cell is a producer cell.

31. A method for producing targeted lipid particles comprising a henipavirus F protein molecule or its biologically active portion, and a targeted envelope protein comprising a henipavirus envelope-binding glycoprotein G (G protein) or its biologically active portion and a single-domain antibody (sdAb) variable domain, a) To provide the cells according to claim 28 or 30, b) Culturing the cells under conditions that enable the generation of targeted lipid particles, and c) Separating, concentrating, or purifying the targeted lipid particles from the cells, thereby producing the targeted lipid particles. The method, including the method described above.

32. A composition comprising a plurality of targeted lipid particles according to any one of claims 1 to 22.

33. The composition according to claim 32, further comprising a pharmaceutically acceptable carrier.

34. Targeted lipid particles according to any one of claims 17 to 22 for use in a method of treating a disease or disorder in a subject, wherein the method comprises delivering an exogenous substance to the subject, and wherein the targeted lipid particles contain the exogenous substance.

35. Use of targeted lipid particles according to any one of claims 17 to 22 in the manufacture of a drug for treating a disease or disorder in a subject.

36. A composition according to claim 32 or 33 for use in a method of treating a disease or disorder in a subject, wherein the method comprises delivering an exogenous substance to the subject, and further wherein the composition comprises a plurality of targeted lipid particles containing the exogenous substance.

37. A targeted lipid particle for use according to claim 34, wherein the exogenous substance is a chimeric antigen receptor (CAR), and the method further comprises delivering the CAR to a target cell, or a composition for use according to claim 36.

38. The targeted lipid particles for use or composition for use according to claim 37, wherein the cells are hepatocytes.

39. Targeted lipid particles according to any one of claims 1 to 22, or a composition according to claim 32 or 33, for use in a method of treating a disease or disorder in a subject, wherein the method comprises fusing mammalian cells to the targeted lipid particles.

40. Targeted lipid particles for use according to any one of claims 34, 37 to 39, wherein the subject is a human subject.

41. The use according to claim 35, wherein the subject is a human subject.

42. The composition for use according to any one of claims 36 to 39, wherein the subject is a human subject.

43. The use according to claim 35, wherein the exogenous substance is a chimeric antigen receptor (CAR), and the treatment comprises delivering the CAR to the target cells.

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