Gene editing compositions of polypeptides and nucleic acids, and their use

Abstract

This specification provides a viral particle comprising a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system, a composition containing the same, and a method of using the same.

Description

[Technical Field]

[0001] Related applications This application claims the interests of U.S. Provisional Patent Application No. 63 / 504,068, filed on 24 May 2023, which is incorporated herein by reference in its entirety.

[0002] Reference to electronically submitted sequence listings This application includes a sequence listing submitted electronically in XML format, the entirety of which is incorporated herein by reference. The XML copy was created on 8 May 2024, is named "INH-022WO_SL", and is 95,139 bytes in size.

[0003] The embodiments provided herein relate to gene-editable polypeptide and nucleic acid compositions, and methods for using them. [Background technology]

[0004] Vesicular stomatitis virus (VSV) is an enveloped minus-strand RNA virus belonging to the genus Vesiculovirus in the family Rhabdoviruses. This virus is an arbovirus that can infect insects, cattle, horses, and pigs. The VSV genome encodes five structural proteins, including a single transmembrane glycoprotein (G). The glycoprotein is a classic type I membrane glycoprotein having an amino-terminal signal peptide, an ectodomain of approximately 450 amino acids, a single alpha-helical transmembrane segment, and a small intraviral carboxy-terminal domain. The signal peptide is cleaved within the lumen of the endoplasmic reticulum, and the native glycoprotein consists of the ectodomain, transmembrane domain, and intraviral domain.

[0005] G plays a crucial role during the initial stages of viral infection (Albertini, AAV, Baquero, E., Ferrin, A., and Gaudin, Y. (2012) Molecular and cellular aspects of rhabdoviral entry. Viruses 4, 117-139.) (This literature is incorporated herein in its entirety by reference). Firstly, it is involved in viral attachment to specific receptors. After binding, virions enter the cell via the clathrin-mediated endocytosis pathway. In the acidic environment of the endocytic vesicle, G induces fusion of the viral membrane with the endosomal membrane, thereby releasing the genome into the cytoplasm for subsequent infection steps. Fusion is catalyzed by a large, low-pH-induced structural transition from the pre-fusion conformation to the post-fusion conformation (both trimers) (Roche, S., Bressanelli, S., Rey, FA, and Gaudin, Y. (2006). Crystal structure of the low-pH form of the vesicular stomatitis virus glycoprotein G. Science 313, 187-191. Roche, S., Rey, FA, Gaudin, Y., and Bressanelli, S. (2007). Structure of the prefusion form of the vesicular stomatitis virus glycoprotein G. Science 315, 843-848) (each of these is incorporated herein by reference in whole).

[0006] The polypeptide chain of the G ectodomain folds into three distinct domains: the fusion domain (FD), the pleckstrin homology domain (PHD), and the trimerization domain (TrD). During the structural transition, FD, PHD, and TrD retain their tertiary structures. Nevertheless, they undergo significant rearrangement in relative orientation due to secondary changes in the hinge segments (S1-S5) that refold during low-pH induced conformational changes (Roche et al., 2006, Roche et al., 2007).

[0007] Low-density lipoprotein receptors (LDL-R) and other members of this receptor family have been shown to function as VSV receptors (Finkelshtein, D., Werman, A., Novick, D., Barak, S., and Rubinstein, M. (2013). LDL receptor and its family members serve as the cellular receptors for vesicular stomatitis virus. Proceedings of the National Academy of Sciences of the United States of America 110, 7306-7311, which is incorporated herein by reference in its entirety). VSV-G can be used to pseudotype other viruses, and VSV-G-pseudotyped lentivirus (VSV-G-LV) exhibits the same broad affinity as VSV. However, this broad affinity may inhibit selective targeting of specific cell types. Therefore, there is a need for a modified (mutant or variant) VSV-G protein that can be used to pseudotype a virus that blocks binding to the LDL receptor. This embodiment satisfies these needs as well as other needs. [Overview of the Initiative]

[0008] In some embodiments, viral particles are provided herein. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the targeting moiety is represented by the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety.

[0009] In some embodiments, the stalk portion S1 comprises a variant Fc protein. In some embodiments, the variant Fc protein comprises a transmembrane domain, such as but not limited to a CD8 or CD28 transmembrane domain. In some embodiments, the variant Fc protein comprises an effector mutation, which inhibits the interaction between the Fc protein and Fc-interacting proteins such as FcγR, C1q, FcRβ, or FcRn.

[0010] In some embodiments, the variant Fc protein is a variant IgG1 Fc protein containing one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A.

[0011] In some embodiments, the variant IgG1 Fc protein contains an amino acid sequence having at least 80% identity to SEQ ID NO: 82, at least 85% identity to SEQ ID NO: 82, at least 90% identity to SEQ ID NO: 82, at least 95% identity to SEQ ID NO: 82, at least 98% identity to SEQ ID NO: 82, or at least 100% identity to SEQ ID NO: 82.

[0012] In some embodiments, the variant Fc protein is a variant IgG2 Fc protein containing one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A.

[0013] In some embodiments, the variant Fc protein is a variant IgG4 Fc protein containing one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A.

[0014] In some embodiments, the targeting moiety having formula T-S1 comprises a stalk moiety S1 having the formula L1-Fc-L2-X1, where L1 is a linker or absent, Fc is a variant Fc protein, L2 is a linker or absent, and X1 is a polypeptide containing a transmembrane domain.

[0015] In some embodiments, the polypeptide containing the transmembrane domain (X1) is ECD-T M - Contains a polypeptide having the formula ICD, where ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent. M ICD is the transmembrane domain of a transmembrane protein, and ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or is absent, and the targeting portion having the formula T-L1-Fc-L2-X1 is T-L1-Fc-L2-ECD-T M - It has the formula ICD.

[0016] In some embodiments, the stalk portion S1 comprises the formula L3-X1, where L3 is a mobile peptide linker and X1 is a polypeptide comprising a transmembrane domain, and the targeting portion having the formula T-S1 comprises the formula T-L3-X1.

[0017] In some embodiments, the polypeptide containing the transmembrane domain (X1) is ECD-T M - Contains a polypeptide having the formula ICD, where ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent. MICD is the transmembrane domain of a transmembrane protein, and ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or is absent, and the targeting portion having the formula T-L3-X1 is T-L3-ECD-T M - It has the formula ICD.

[0018] In some embodiments, the targeted portion binds to CD7. In some embodiments, the targeted portion comprises a polypeptide comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 30, the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 31, and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 32, or any of the aforementioned variants; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 33, the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 34, and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 35, or any of the aforementioned variants provided herein.

[0019] In some embodiments, the targeting portion binds to CD8. In some embodiments, the targeting portion comprises a polypeptide comprising: (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 42, the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 43, and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 44, or any of the aforementioned variants; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 45, the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 46, and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 47, or any of the aforementioned variants provided herein.

[0020] In some embodiments, the heterologous viral glycoprotein is an SVCV-G polypeptide. In some embodiments, the SVCV-G polypeptide is as provided herein.

[0021] In some embodiments, the heterologous viral glycoprotein is a VSV-G polypeptide. In some embodiments, the VSV-G polypeptide is as provided herein. In some embodiments, the VSV-G polypeptide includes substitutions at positions I182, T214, and T352 of SEQ ID NO: 2. In some embodiments, the substitution at position I82 is I182D or I182E. In some embodiments, the substitution at position 214 is T214N. In some embodiments, the substitution at position 352 is T352A.

[0022] In some embodiments, the gene editing system is a CRISPR-Cas system, a zinc finger nuclease system, a TALEN, a meganuclease, or a gene product regulatory nucleic acid molecule. In some embodiments, the nucleic acid molecule encodes a CRISPR-Cas system comprising a Cas protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or a guide RNA. In some embodiments, the gene product regulatory nucleic acid molecule encodes at least one molecule selected from siRNA, piRNA, miRNA, RNAi RNA, mRNA, shRNA, and antisense RNA.

[0023] In some embodiments, methods for infecting cells are provided. In some embodiments, the method includes bringing cells into contact with viral particles provided herein.

[0024] In some embodiments, a method is provided for infecting cells in a subject. In some embodiments, the method includes administering to the subject a pharmaceutical composition comprising viral particles provided herein.

[0025] In some embodiments, methods are provided for editing a target nucleic acid molecule within a cell. In some embodiments, the method involves contacting a cell with a viral particle provided herein, wherein a nucleic acid molecule encoding a gene editing system is expressed within the cell to edit the target nucleic acid molecule within the cell.

[0026] In some embodiments, methods are provided for treating a disease or disorder in a subject. In some embodiments, the method comprises administering viral particles provided herein to a subject, wherein a nucleic acid molecule encoding a gene editing system is expressed in the subject, and the subject is treated for a disease or disorder.

[0027] In some embodiments, a viral particle is provided. In some embodiments, the viral particle includes (a) a VSV-G polypeptide having an amino acid sequence having at least 70% identity with SEQ ID NO: 2 and having a mutation at position 182 compared to SEQ ID NO: 2; (b) a nucleic acid molecule encoding a gene editing system, wherein the gene editing system is a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets a guide RNA, a single guide RNA, or a Cas protein to a target nucleic acid molecule; (c) optionally, a nucleic acid molecule encoding a heterologous molecule of interest; and (d) a targeting moiety that binds to a T cell, CD3+ T cell, CD4+ T cell, CD7+ T cell, or CD8+ T cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0028] In some embodiments, a virus particle is provided. In some embodiments, the virus particle comprises (a) an SVCV-G polypeptide having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with respect to SEQ ID NO: 52 or SEQ ID NO: 53, and (b ) A nucleic acid molecule encoding a gene editing system, wherein the gene editing system is a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets a target nucleic acid molecule, a guide RNA, a single guide RNA, or a Cas protein; (c) optionally, a nucleic acid molecule encoding a heterologous molecule of interest; and (d) a targeting moiety that binds to a T cell, CD3+ T cell, CD4+ T cell, CD7+ T cell, or CD8+ T cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0029] In some embodiments, a method for editing a target nucleic acid within a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle, the viral particle comprising: (a) a VSV-G polypeptide having at least 70% identity to SEQ ID NO: 2 and having a mutation at position 182 compared to SEQ ID NO: 2; (b) a nucleic acid molecule encoding a gene editing system, wherein the gene editing system is a CRISPR-Cas system comprising a Cas9 protein and guide RNA, single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule; (c) optionally, a nucleic acid molecule encoding a heterologous molecule of interest; and (d) a targeting moiety that binds to a T cell, CD3+ T cell, CD4+ T cell, CD7+ T cell, or CD8+ T cell, wherein the nucleic acid molecule encoding the gene editing system is expressed within the cell to edit the target nucleic acid molecule within the cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0030] In some embodiments, a method for editing a target nucleic acid within a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle, the viral particle comprising (a) an SVCV-G polypeptide having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 52 or SEQ ID NO: 53, and (b) a gene editing system. A nucleic acid molecule encoding a gene editing system which is a CRISPR-Cas system comprising a Cas9 protein and guide RNA, single guide RNA, or at least one nucleic acid molecule that targets a target nucleic acid molecule with a Cas protein; (c) optionally a nucleic acid molecule encoding a heterologous molecule of interest; and (d) a targeting portion that binds to T cells, CD3+ T cells, CD4+ T cells, CD7+ T cells, or CD8+ T cells, wherein the nucleic acid molecule encoding the gene editing system is expressed in a cell and edits the target nucleic acid molecule in the cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0031] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stalk moiety S1 comprises a variant Fc protein comprising an amino acid sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28. In some embodiments, the variant of SEQ ID NO: 26 comprises one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant of SEQ ID NO: 28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant Fc protein further comprises a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a Cas protein.

[0032] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stalk portion (S1) comprises the formula -L1-Fc-L2-X1, where L1 is a linker containing the sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, or SEQ ID NO: 76, or is absent, and Fc is a variant Fc protein containing a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28, where the variant of SEQ ID NO: 26 is selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. The variant of SEQ ID NO: 27 contains one or more mutations, the variant of SEQ ID NO: 27 contains one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A, the variant of SEQ ID NO: 28 contains one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A, L2 is a linker containing the sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, or SEQ ID NO: 76, or is not present, and X1 is ECD-T M - A polypeptide comprising a transmembrane domain having the formula ICD. In some embodiments, ECD is an extracellular domain having the sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent. In some embodiments, T MThis is a transmembrane domain having the sequence of SEQ ID NO: 61 or SEQ ID NO: 62, or a fragment thereof. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and the ICD includes an env incorporation motif having the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a guide RNA.

[0033] In some embodiments, virus particles are provided. In some embodiments, the virus particles comprise a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk portion. In some embodiments, the target binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stalk portion (S1) comprises the formula -L1-Fc-L2-X1, wherein L1 is a linker comprising the sequence of SEQ ID NO: 55 or is absent, Fc is a variant Fc protein comprising a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28, the variant of SEQ ID NO: 26 comprises one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A, the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A, the variant of SEQ ID NO: 28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A, L2 is a linker comprising the sequence of SEQ ID NO: 55 or is absent, and X1 is ECD-T M -a polypeptide comprising a transmembrane domain having the formula ICD. In some embodiments, the ECD is an extracellular domain having the sequence of SEQ ID NO: 60 or a fragment thereof or is absent. In some embodiments, T MThis is a transmembrane domain having the sequence of SEQ ID NO: 62, or a fragment thereof. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD contains an env incorporation motif having the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or Cas protein.

[0034] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral glycoprotein comprises the sequence of SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stalk moiety S1 comprises the formula L1-Fc-L2-X1, where L1 is a linker comprising the sequence of SEQ ID NO: 55, Fc is a variant Fc protein comprising the sequence of SEQ ID NO: 82, L2 is a linker and is absent, and X1 is ECD-T M - A polypeptide comprising a transmembrane domain having the formula ICD. In some embodiments, ECD is an extracellular domain having the sequence of SEQ ID NO: 60. In some embodiments, T MThis is a transmembrane domain having the sequence of SEQ ID NO: 62. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD includes an env incorporation motif having the amino acid sequence of SEQ ID NO: 63. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a guide RNA.

[0035] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral glycoprotein comprises a sequence having at least 90% identity to SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identity to SEQ ID NO: 23 or SEQ ID NO: 25, at least 98% identity to SEQ ID NO: 23 or SEQ ID NO: 25, or at least 90% identity to SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the targeting moiety comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 83, at least 95% identity to SEQ ID NO: 83, at least 98% identity to SEQ ID NO: 83, or at least 100% identity to SEQ ID NO: 83. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, such as a guide RNA or single guide RNA.

[0036] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral glycoprotein comprises a sequence having at least 90% identity to SEQ ID NO: 52 or SEQ ID NO: 53, at least 95% identity to SEQ ID NO: 52 or SEQ ID NO: 53, at least 98% identity to SEQ ID NO: 52 or SEQ ID NO: 53, or at least 90% identity to SEQ ID NO: 52 or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 83, at least 95% identity to SEQ ID NO: 83, at least 98% identity to SEQ ID NO: 83, or at least 100% identity to SEQ ID NO: 83. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, such as a guide RNA, a single guide RNA, or a Cas protein.

[0037] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38 or SEQ ID NO: 39. In some embodiments, the stalk moiety S1 comprises the formula L3-X1, where L3 is a mobile linker comprising the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, and X1 is the formula ECD-T M- A polypeptide comprising a transmembrane domain having an ICD. In some embodiments, the ECD is an extracellular domain having the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent. In some embodiments, T M This is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD may or may not contain an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a Cas protein.

[0038] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 50 or SEQ ID NO: 51. In some embodiments, the stalk moiety S1 comprises the formula L3-X1, where L3 is a mobile linker comprising the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, and X1 is a molecule of the formula ECD-T M - A polypeptide comprising a transmembrane domain having an ICD. In some embodiments, the ECD is an extracellular domain having the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent. In some embodiments, T MThis is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD may or may not contain an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a Cas protein.

[0039] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral structural protein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stalk moiety S1 comprises the formula L3-X1, where L3 is a mobile linker comprising the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, and X1 is the formula ECD-T M - A polypeptide comprising a transmembrane domain having an ICD. In some embodiments, the ECD is an extracellular domain having the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent. In some embodiments, T MThis is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, or a fragment thereof. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD contains or does not contain an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or a Cas protein.

[0040] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral structural protein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stalk moiety S1 comprises the formula L3-X1, where L3 is a mobile linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a linker of the formula ECD-T M - A polypeptide comprising a transmembrane domain having an ICD. In some embodiments, the ECD is an extracellular domain comprising the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, T MThis is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61, or a fragment thereof. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD includes an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a guide RNA.

[0041] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral structural protein comprises the sequence of SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stalk moiety S1 comprises the formula L3-X1, where L3 is a mobile linker comprising the amino acid sequence of SEQ ID NO: 55 and X1 is a molecule of the formula ECD-T M - A polypeptide comprising a transmembrane domain having an ICD. In some embodiments, the ECD is an extracellular domain comprising the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, T MThis is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61, or a fragment thereof. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD includes an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a guide RNA.

[0042] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the heterologous viral structural protein comprises the sequence of SEQ ID NO: 52 or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stalk moiety S1 comprises the formula L3-X1, where L3 is a mobile linker comprising the amino acid sequence of SEQ ID NO: 55 and X1 is of the formula ECD-T M - A polypeptide comprising a transmembrane domain having an ICD. In some embodiments, the ECD is an extracellular domain comprising the amino acid sequence of SEQ ID NO: 59, or a fragment thereof. In some embodiments, T MThis is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61, or a fragment thereof. In some embodiments, the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD includes an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, single guide RNA, or a guide RNA. [Brief explanation of the drawing]

[0043] [Figure 1A] The crystal structures of VSV-G bound to LDL-R are shown. Figure 1A shows the crystal structure of VSV-G bound to CR3 of LDL-R. Figure 1B shows the crystal structure of VSV-G bound to CR2 of LDL-R. [Figure 1B] The crystal structures of VSV-G bound to LDL-R are shown. Figure 1A shows the crystal structure of VSV-G bound to CR3 of LDL-R. Figure 1B shows the crystal structure of VSV-G bound to CR2 of LDL-R.

[0044] [Figure 2A] This shows the effect of adding negatively charged amino acids to VSV-G. The LDL-R bond interacts with innate affinity and membrane fusion properties. Figure 2A shows the titration of VSV-G constructs on SupT1 cells. Figure 2B shows the functional titer of each construct calculated from the titration in Figure 2A. [Figure 2B] This shows the effect of adding negatively charged amino acids to VSV-G. The LDL-R bond interacts with innate affinity and membrane fusion properties. Figure 2A shows the titration of VSV-G constructs on SupT1 cells. Figure 2B shows the functional titer of each construct calculated from the titration in Figure 2A.

[0045] [Figure 3-1]This shows the alignment of ectodomains of different VSV-G proteins from different strains. [Figure 3-2] This shows the alignment of ectodomains of different VSV-G proteins from different strains. [Figure 3-3] This shows the alignment of ectodomains of different VSV-G proteins from different strains. [Figure 3-4] This shows the alignment of ectodomains of different VSV-G proteins from different strains.

[0046] [Figure 4] This study demonstrates the effects of various VSV-G mutations on the serum stability of viral constructs.

[0047] [Figure 5] This study demonstrates the effects of various VSV-G mutations on the serum stability of viral constructs combined with a CD7 binder.

[0048] [Figure 6] The ability of various rhabdoviral G proteins to transduce SupT1 and PBMC cells, either alone or in combination with a CD7 binder, is demonstrated.

[0049] [Figure 7A] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7B] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7C] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7D] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7E]Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7F] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7G] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7H] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7I] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7J] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7K] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 7L] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0050] [Figure 7M] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0051] [Figure 8A] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8B]Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8C] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8D] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8E] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8F] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8G] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8H] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8I] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8J] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8K] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8L] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0052] [Figure 8M] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0053] [Figure 9A] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD8 binder disclosed herein are shown.

[0054] [Figure 9B] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD8 binder disclosed herein are shown.

[0055] [Figure 10A] VSV-G* pseudotyped lentivirus particles containing a CD7 binder with mutant Fc stalks demonstrate the ability to transduce SupT1 cells as well as human and non-human primate PBMCs. [Figure 10B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 10C] The transduction of human and non-human primate PBMCs is shown from the perspective of MOI calculated from SupT1 titration. VSV-G* refers to VSV-G (I182E, T214N, T352A).

[0056] [Figure 11A] SVCV-G pseudotyped lentiviral particles containing a CD7 binder with mutant Fc stalks demonstrate the ability to transduce SupT1 cells and human and non-human primate PBMCs. [Figure 11B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 11C] From the perspective of MOI calculated from SupT1 titration, we demonstrate the transduction of human and non-human primate PBMCs.

[0057] [Figure 12A]This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 12B] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 12C] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 12D] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A).

[0058] [Figure 13A] This is a comparison of off-target GFP transduction in a panel of B cell lines compared to control SupT1 cells. Figure 13A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 13B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks. [Figure 13B]This is a comparison of off-target GFP transduction in a panel of B cell lines compared to control SupT1 cells. Figure 13A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 13B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks.

[0059] [Figure 14A] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to control SupT1 cells. Figure 14A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 14B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks. [Figure 14B] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to control SupT1 cells. Figure 14A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 14B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with mutant Fc stalks.

[0060] [Figure 15A] VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk demonstrate the ability to transduce SupT1 cells and human and non-human primate PBMCs. [Figure 15B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 15C]The transduction of human and non-human primate PBMCs is shown from the perspective of MOI calculated from SupT1 titration. VSV-G* refers to VSV-G (I182E, T214N, T352A).

[0061] [Figure 16A] This study demonstrates the ability of viral particles containing CD7 binders with mobile stalks of varying lengths to transduce SupT1 cells, compared to other IgG-based binders. [Figure 16B] This study demonstrates the ability of viral particles containing CD7 binders with mobile stalks of varying lengths to transduce PBMC cells, compared to other IgG-based binders.

[0062] [Figure 17A] SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk demonstrate the ability to transduce SupT1 cells and human and non-human primate PBMCs. [Figure 17B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 17C] From the perspective of MOI calculated from SupT1 titration, we demonstrate the transduction of human and non-human primate PBMCs.

[0063] [Figure 18A] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 18B] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 18C]This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 18D] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A).

[0064] [Figure 19A] This is a comparison of off-target GFP transduction in a panel of B cell lines compared to the control SupT1. Figure 19A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 19B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk. [Figure 19B] This is a comparison of off-target GFP transduction in a panel of B cell lines compared to the control SupT1. Figure 19A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 19B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk.

[0065] [Figure 20A]This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to the control SupT1. Figure 20A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 20B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk. [Figure 20B] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to the control SupT1. Figure 20A shows data for VSV-G* pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). Figure 20B shows data for SVCV-G pseudotyped lentivirus particles possessing a CD7 binder with a mobile stalk.

[0066] [Figure 21A] Figure 21A shows the reduction in the percentage of cells positive for the target gene after transduction with the lentiviral constructs of this disclosure. Figure 21A shows the reduction in GFP-positive cells after treatment with lentiviral constructs containing Cas9 and GFP sgRNA. No reduction in GFP-positive cells is observed when the lentiviral construct delivers CD7 sgRNA. Figure 21B shows the reduction in CD7-positive cells after treatment with lentiviral constructs containing Cas9 and one of two independent CD7 sgRNAs. No reduction in CD7-positive cells is observed when the lentiviral construct delivers GFP sgRNA. Data are from 7 days after transduction with 100 μL of virus. [Figure 21B]Figure 21A shows the reduction in the percentage of cells positive for the target gene after transduction with the lentiviral constructs of this disclosure. Figure 21A shows the reduction in GFP-positive cells after treatment with lentiviral constructs containing Cas9 and GFP sgRNA. No reduction in GFP-positive cells is observed when the lentiviral construct delivers CD7 sgRNA. Figure 21B shows the reduction in CD7-positive cells after treatment with lentiviral constructs containing Cas9 and one of two independent CD7 sgRNAs. No reduction in CD7-positive cells is observed when the lentiviral construct delivers GFP sgRNA. Data are from 7 days after transduction with 100 μL of virus.

[0067] [Figure 22A] Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22B]Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22C] Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22D]Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22E] Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22F]Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22G] Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22H]Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22I] Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22J]Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22K] Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L). [Figure 22L]Figures 22A to 22D show the presence of distinct GFP-negative or CD7-negative cell populations 19 days after transduction. Samples treated with 100 μL of CD7 sgRNA-containing virus (Figures 22A and 22B) or GFP sgRNA-containing virus (Figure 22C) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22D). Figures 22E to 22H show samples treated with 20 μL of CD7 sgRNA-containing virus (Figures 22E and 22F) or GFP sgRNA-containing virus (Figure 22G) are compared with untreated parental GFP-positive and CD7-positive cells (Figure 22H). Figures 22I to 22L compare samples treated with 4 μL of CD7 sgRNA-containing virus (Figures 22I and 22J) or GFP sgRNA-containing virus (Figure 22K) with untreated parental GFP-positive and CD7-positive cells (Figure 22L).

[0068] [Figure 23] This shows GFP expression in cells treated with CD7 sgRNA-containing virus or GFP sgRNA-containing virus, compared to parental GFP-positive and CD7-positive cells. Neither CD7 sgRNA construct resulted in a decrease in GFP expression. However, the GFP sgRNA construct delivered with the viral constructs of this disclosure resulted in a robust and observable decrease in GFP expression. Wells marked with an asterisk indicate that these cells were harvested for further proliferation.

[0069] [Figure 24] This shows the results of genomic DNA sequence analysis around the expected Cas9 cleavage site in cells transduced with GFP sgRNA. The analysis indicates that the GFP gene is cleaved at the expected cleavage site determined by the guide sequence.

[0070] [Figure 25] This shows the results of genomic DNA sequence analysis around the expected Cas9 cleavage site in cells transduced with CD7_3 sgRNA. The analysis indicates that the CD7 gene is cleaved at the expected cleavage site determined by the guide sequence. [Modes for carrying out the invention]

[0071] This specification provides a viral particle that may comprise, for example, a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. The targeting moiety may be in the form of T-L1-Fc-L2-X1 or T-L3-X1, where T is the targeting moiety, L1 is a linker or absent, Fc is a mutant Fc protein, L2 is a linker or absent, L3 is a mobile peptide linker, and X1 is a polypeptide comprising a transmembrane domain. The mutant Fc polypeptide may be incorporated into the viral particle to help facilitate the targeting of the viral particle to a particular cell type. The at least one nucleic acid molecule encoding a gene editing system may encode a CRISPR-Cas system comprising a gene editing system, for example, a Cas protein, and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, such as a guide RNA, a single guide RNA, or a guide RNA.

[0072] Furthermore, the viral particles may contain a VSV-G protein, which can be used to pseudotype viruses, such as lentiviruses. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of the New Jersey virus strain for bullous stomatitis, the Indiana virus strain for bullous stomatitis, the Aragoas virus strain for bullous stomatitis, the Maraba virus strain for bullous stomatitis, or the Karajas virus strain for bullous stomatitis. Examples of such proteins are provided herein.

[0073] Pseudotyped viruses containing a variant VSV-G protein, for example, those provided herein, can be used in conjunction with a targeting moiety to facilitate fusion of the pseudotyped virus with specific cells or tissues based on the expression of a target on the cell or tissue. As provided herein, the targeting moiety may be linked to an Fc protein, which may be called a stalk protein, containing a transmembrane domain to facilitate the attachment of the targeting moiety to the surface of the virus. In some embodiments, the Fc protein includes an Fc effector mutation, such as those provided herein. As provided herein, the targeting moiety may alternatively be linked to a mobile polypeptide, which may be called a "mobile stalk protein" or "mobile stalk," containing a transmembrane domain to facilitate the attachment of the targeting moiety to the surface of the virus. In some embodiments, the mobile polypeptide is a mobile peptide, for example, those provided herein.

[0074] Unless otherwise defined, all technical and scientific terms have the same meaning as they would be generally understood by those skilled in the art in the field to which the disclosed embodiments belong.

[0075] As used herein, the terms "a" or "an" mean "at least one" or "one or more" unless the context clearly indicates otherwise.

[0076] Where used herein, the term “about” means that a number is an approximation and small variations will not significantly affect the implementation of the disclosed embodiment. Where numerical limitations are used, unless otherwise indicated by the context, “about” means that the number may vary by ±10% and remain within the range of the disclosed embodiment. Furthermore, where the phrase “about x to y” is used, the term “about” can be used interchangeably with the phrase “about x to y” unless both x and y are modified to indicate a different context.

[0077] As used herein, the terms “individual,” “subject,” or “patient” are used synonymously and mean any animal, including mice, rats and other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates such as humans.

[0078] As used herein, the terms “comprising” (and any conjugations of “comprising,” such as “comprise,” “comprises,” and “comprised,” “having” (and any conjugations of “having,” such as “have,” and “has,” “has,” “having,” “having,” “having,” “having,” “having,” “having,” “having,” “having,” and any conjugations of “having,” such as “havings,” and “having,” or “having,” “having,” are open or non-restrictive and do not exclude any additional elements or steps of method not described herein. Any step or composition using the transitional phrase “comprise” or “comprising” can also be said to describe the same thing using the transitional phrase “consisting of” or “consists.”

[0079] As used herein, the term “contact” means bringing two elements together in vitro or in vivo. For example, “contact” a virus or vector described herein with an individual, patient, or cells includes administering the virus to an individual or patient, such as a human, and introducing a compound into a sample containing, for example, a cell preparation or purified preparation containing such cells.

[0080] As used herein, the terms “fused” or “linked” used in reference to proteins having different domains or heterologous sequences mean that such protein domains are connected to each other by either peptide bonds or other covalent bonds and become part of the same peptide chain. Domains or sections may be directly linked or fused to each other, or another domain or peptide sequence may be between two domains or sequences, and such sequences will still be considered fused or linked to each other. In some embodiments, the various domains or proteins provided herein may be directly linked or fused to each other, or a linker sequence, such as the glycine / serine sequence described herein, may link the two domains together.

[0081] As used herein, “mobile stalk” or “mobile stalk protein” refers to a polypeptide containing a mobile region, which in some embodiments may be ligated to an extracellular domain, a transmembrane domain, and / or an intracellular domain. The extracellular domain may be a target-binding domain, which may be denoted as “T”.

[0082] In some embodiments, “mutant” or “variant” Fc proteins are provided. As used herein, “mutant” (or “mutated”) and “variant” are used interchangeably to indicate that the provided Fc protein contains one or more mutations compared to the natural or wild-type Fc protein. Therefore, it should be understood that the terms “mutant Fc” and “variant Fc” are considered synonymous unless the context requires otherwise.

[0083] A "disease" is a state of animal health in which the animal is unable to maintain homeostasis, and if the disease is not improved, the animal's health will continue to deteriorate. In contrast, a "disorder" in animals is a state of health in which the animal can maintain homeostasis, but the animal's health is less desirable than when the disorder is absent. If left untreated, a disorder does not necessarily lead to a further decline in the animal's health.

[0084] "Effective dose" or "therapeutic effective dose" is used synonymously herein and refers to the amount of a compound, formulation, substance, or composition described herein that is effective in achieving a particular biological outcome or in producing a therapeutic or preventive effect. Such outcomes may include, but are not limited to, an amount that causes a detectable level of immune cell activation when administered to a mammal, compared to immune cell activation detected in the absence of the composition. The immune response can be readily assessed by many methods recognized in the art. Those skilled in the art will understand that the amounts of compositions administered herein vary and can be readily determined based on several factors, such as the disease or condition being treated, the age, health status, and physical condition of the mammal being treated, the severity of the disease, and the specific compound being administered.

[0085] "Code" refers to the inherent property of a particular sequence of nucleotides in a polynucleotide (e.g., a gene, cDNA, or mRNA) to function as a template for the synthesis of other polymers and macromolecules having either a specific sequence of nucleotides (i.e., rRNA, tRNA, and mRNA) or a specific sequence of amino acids in a biological process, as well as the biological properties derived therefrom. Therefore, a gene codes for a protein when the transcription and translation of the mRNA corresponding to that gene produces a protein in a cell or other biological system. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in a sequence listing, and the non-coding strand, used as a template for the transcription of a gene or cDNA, can be said to code for a protein or other product of that gene or cDNA.

[0086] An "expression vector" refers to a vector containing recombinant polynucleotides that include an expression control sequence operably ligated to the nucleotide sequence to be expressed. An expression vector contains sufficient cis-acting elements for expression, and other elements for expression may be supplied by a host cell or in an in vitro expression system. Expression vectors include all known in the art, such as cosmids, plasmids (e.g., naked or liposome-containing) and viruses (e.g., Sendai virus, lentivirus, retrovirus, adenovirus, and adeno-associated virus) that incorporate recombinant polynucleotides.

[0087] As used herein, the term “ex vivo” in relation to cells transfected, transfected, or transformed ex vivo means cells that are transfected, transfected, or transformed outside of the subject, i.e., cells that are removed from the subject before such cells are transfected, transfected, or transformed.

[0088] As used herein, “identity” refers to the identity of subunit sequences between two nucleic acid molecules or amino acid molecules, or between two polymer molecules, such as two polynucleotide molecules or polypeptide molecules. Two amino acid sequences are identical at the same position if, for example, the position in each of two polypeptide molecules is occupied by arginine. The identity or degree to which two amino acids or two nucleic acid sequences have the same residue at the same position in their alignment is often expressed as a percentage. The identity between two amino acids or two nucleic acid sequences is a linear function of the number of matching or identical positions; for example, if half of the positions in the two sequences are identical, the two sequences are 50% identical, and if 90% of the positions (e.g., 9 out of 10) match or are identical, the two amino acid sequences are 90% identical.

[0089] "Substantially identical" means that a polypeptide or nucleic acid molecule exhibits at least 50% identity with respect to a reference amino acid sequence (e.g., any one of the amino acid sequences described herein) or nucleic acid sequence (e.g., any one of the nucleic acid sequences described herein). In some embodiments, such sequences are at least 60%, 80%, or 85%, or 90%, 95%, or even 99%, identical at the amino acid level or nucleic acid level to a sequence used for comparison. Other identity percentages with respect to specific sequences are described herein.

[0090] Sequence identity can be measured / determined using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX program). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine, valine, isoleucine, leucine, aspartic acid, glutamic acid, asparagine, glutamine, serine, threonine, lysine, arginine, and phenylalanine, tyrosine. An exemplary method for determining the degree of identity can be used with the BLAST program, where probability scores from e3 to e100 indicate closely related sequences. In some embodiments, sequence identity is determined by using BLAST with default settings.

[0091] The scope of embodiments provided herein includes compositions comprising various proteins, which may, in some cases, include amino acid sequences having sequence identity with the amino acid sequences disclosed herein. Therefore, in certain embodiments, depending on the specific sequence, the degree of sequence identity is preferably greater than 50% with respect to the SEQ ID NOs disclosed herein (e.g., 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more). In addition to these percentages, other identity percentages are also indicated herein. Identity between polypeptides can be determined by a Smith-Waterman homology search algorithm, such as that implemented in the MPSRCH program (Oxford Molecular), using an affine gap search with a gap-opening penalty of 12 and a gap-extending penalty of 1.

[0092] These proteins may contain one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conserved amino acid substitutions, i.e., substitutions of one amino acid with another amino acid having a related side chain, compared to the disclosed proteins. Genetically encoded amino acids are generally divided into four families: (1) acidic, i.e., aspartic acid, glutamic acid; (2) basic, i.e., lysine, arginine, histidine; (3) nonpolar, i.e., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) non-charged, i.e., glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes classified together as aromatic amino acids. In general, single amino acid substitutions within these families do not have a significant impact on biological activity. The protein may have one or more single amino acid deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) compared to the disclosed protein sequence. The protein may also have one or more insertions (e.g., each of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) (e.g., each of 1, 2, 3, 4, or 5 amino acids) compared to the disclosed protein sequence.

[0093] As used herein, the term "in vivo" with respect to cells transfected, transformed, or otherwise transfected in vivo means that the cells are transfected, transformed, or otherwise transfected within the subject, and that the cells have not been removed from the subject before being transfected, transfected, or transformed.

[0094] "Isolated" means modified or extracted from its natural state. For example, nucleic acids or peptides that are naturally present in living animals are not "isolated," but the same nucleic acids or peptides that have been partially or completely separated from their naturally occurring coexisting materials are "isolated." Isolated nucleic acids or proteins can exist in a substantially purified form or in a non-natural environment, such as a host cell.

[0095] As used herein, "lentivirus" refers to a genus of retroviridae capable of infecting non-dividing cells. Non-exclusive examples of lentiviruses include HIV, SIV, and FIV. Lentivirus-derived vectors or virus-like particles can be used to transduce cells, deliver genes or other molecules, and express them intracellularly, either in vitro (ex vivo) or in vivo.

[0096] The term "modified," as used herein, means an altered state or structure of a molecule or cell provided herein. Molecules can be modified in many ways, including chemical, structural, and functional modifications such as mutation, substitution, insertion, or deletion (e.g., internal deletion cleavage). Cells can be modified by the introduction of nucleic acids or the expression of heterologous proteins.

[0097] As used herein, the term “modulation” means mediating an increase or decrease in the response level of a subject compared to the response level of the subject in the absence of the treatment or compound, and / or compared to the response level of an otherwise identical but untreated subject. This term includes mediating a beneficial therapeutic response in a subject such as a human by disrupting and / or influencing a natural signal or response.

[0098] Unless otherwise specified, "nucleotide sequences encoding an amino acid sequence" includes all nucleotide sequences that encode the same amino acid sequence, including degenerate versions of each other. The phrase "nucleotide sequence encoding a protein or RNA" may include introns to the extent that a nucleotide sequence encoding a protein may contain introns in several versions.

[0099] The term "oligonucleotide" typically refers to a short polynucleotide. If a nucleotide sequence is represented by a DNA sequence (i.e., A, T, C, G), it is understood that this also provides a corresponding RNA sequence (i.e., A, U, C, G) where "T" is replaced by "U".

[0100] Parenteral administration of the composition includes, for example, subcutaneous (sc), intravenous (iv), intramuscular (im), or intrasternal injection, or infusion techniques.

[0101] As used herein, the term “polynucleotide” is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Therefore, as used herein, the terms “nucleic acid” and “polynucleotide” are interchangeable. As used herein, polynucleotide includes, but is not limited to, all nucleic acid sequences obtained by any method available in the art, including, but not limited to, recombinant methods such as cloning of nucleic acid sequences from recombinant libraries or cell genomes using cloning techniques and PCR, etc.

[0102] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used synonymously and refer to compounds composed of multiple amino acid residues covalently linked by peptide bonds. As used herein, this term refers to both short chains, also commonly referred to in the art as peptides, oligopeptides, and oligomers, and long chains, of which there are many types, commonly referred to in the art as proteins. Polypeptides include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, polypeptide variants, modified polypeptides, derivatives, analogs, and fusion proteins. Polypeptides include native peptides, recombinant peptides, synthetic peptides, or combinations thereof.

[0103] The terms “pseudotyped” or “pseudotyped viral particle,” as used herein, refer to viral particles having glycoproteins derived from another virus that has an envelope or envelope glycoprotein encoding a viral vector from a virus different from the parent virus. Thus, the host range of the vector particle can be expanded or modified depending on the type of cell surface receptor used by the glycoprotein. For example, a virus may be pseudotyped with a VSV-G mutant protein, as provided herein.

[0104] Where used herein in relation to antibodies, the term “specifically binds” means that an antibody recognizes a particular antigen while substantially not recognizing or binding to other molecules in the sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. However, such interspecies reactivity does not in itself alter the antibody’s classification as specific. In another example, an antibody that specifically binds to an antigen may also bind to different alleles of the antigen. However, such cross-reactivity does not in itself alter the antibody’s classification as specific. In some cases, the term “specifically binds” or “specifically binds” can be used in relation to the interaction of an antibody, protein, or peptide with a second chemical species, meaning that the interaction depends on the presence of a specific structure on the chemical species (e.g., an antigenic determinant or epitope), for example, that an antibody recognizes and binds to a specific protein structure rather than to proteins in general. If an antibody is specific to epitope “A”, then in a reaction involving labeled “A” and the antibody, the presence of a molecule containing epitope A (or unlabeled free A) will reduce the amount of labeled A that binds to the antibody. In some embodiments, the targeting moieties described herein can be used to target viral particles containing a variant VSV-G protein, or to target viral particles containing other viral structural proteins used to pseudotype the virus, such targeting moieties can specifically bind to their target.

[0105] The term "subject" includes living organisms, including organisms (e.g., mammals) from which an immune response can be induced. As used herein, "subject" or "patient" may be human or a non-human mammal. Non-human mammals include, for example, livestock and pets such as sheep, cattle, pigs, dogs, non-human primates, cats, and mouse mammals. In some embodiments, the subject is human.

[0106] As used herein, the term “therapeutic” means treatment and / or prevention. Therapeutic effects are obtained by suppression, remission, or eradication of a disease state.

[0107] As used herein, the terms “transfected,” “transformed,” or “transduced” refer to the process by which an exogenous nucleic acid is transferred to or introduced into a cell. A “transfected,” “transformed,” or “transduced” cell is a cell that has been transfected, transformed, or transduced with an exogenous nucleic acid. Cells include primary target cells and their offspring. In some embodiments, transfection, transformation, or transduction is performed or carried out in vivo.

[0108] As used herein, “treating” a disease means reducing the frequency or severity of at least one sign or symptom of the disease or disorder experienced by the subject.

[0109] A "vector" is a composition of substances containing isolated nucleic acids that encode a protein or peptide. Numerous vectors are known in the art, including, but not limited to, linear polynucleotides, plasmids, DNA, and RNA. Examples of viral vectors include, but are not limited to, Sendai virus vectors, adenovirus vectors, adeno-associated virus vectors, retrovirus vectors, and lentiviral vectors.

[0110] The "carrier" or "delivery vehicle" comprises viral particles, viruses, polylysine compounds, and liposomes, which facilitate the transfer of nucleic acids into cells. The carrier or delivery vehicle can also be used to deliver proteins or peptides to cells.

[0111] Scope: Throughout this disclosure, various aspects of the embodiments can be expressed in the form of a range. It should be understood that the use of range format is merely for convenience and conciseness and should not be interpreted as a definitive limitation. Therefore, a range should be considered to specifically disclose all possible subranges, as well as the individual numbers within that range. For example, a range such as 1 to 6 should be considered to specifically disclose subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, and the individual numbers within that range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the width of the range. Unless otherwise explicitly stated, the disclosed range includes the endpoints of that range.

[0112] Virus particles In some embodiments, a viral particle is provided that comprises a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system.

[0113] In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is the target-binding domain and S1 is the stalk moiety. In some embodiments, S1 comprises a variant Fc protein, which comprises a transmembrane domain such as, but not limited to, a CD8 or CD28 transmembrane domain. Thus, in some embodiments, the stalk moiety S1 comprises an N-terminal to C-terminal orientation of the variant Fc transmembrane domain. In some embodiments, the variant Fc protein comprises an effector mutation, which inhibits the interaction between the Fc protein and Fc-interacting proteins such as FcγR, C1q, FcRβ, or FcRn.

[0114] In some embodiments, the S1 stalk portion is attached to the surface of the viral particle via a transmembrane domain. In some embodiments, the Fc protein is an IgG1 Fc, IgG2 Fc, or IgG4 Fc protein. In some embodiments, the mutant Fc protein includes a variant of the sequence of SEQ ID NO: 26 (IgG1 Fc), SEQ ID NO: 27 (IgG2 Fc), or SEQ ID NO: 28 (IgG4 Fc).

[0115] In some embodiments, the variant Fc protein is the variant IgG1 Fc protein (SEQ ID NO: 26). In some embodiments, the variant IgG1 Fc protein contains one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26, numbered according to Kabat's EU numbering index, as described in Edelman, GM et al. "The covalent structure of an entire gammaG immunoglobulin molecule." Proceedings of the National Academy of Sciences of the United States of America vol.63,1(1969):78-85.doi:10.1073 / pnas.63.1.78 (which is incorporated herein by reference in its entirety). Any of the mutations L234A, L235A, N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 26 may or may not be present, and these mutations may be combined in any combination. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A and L235A in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to N297A in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to P329G in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, and P329G in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to I253A in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H310A in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H435A in SEQ ID NO: 26.In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to I253A, H310A, and H435A in SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 26.

[0116] In some embodiments, the variant Fc protein, which includes the variant IgG1 Fc protein, includes a cleavage of the IgG1 Fc sequence. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the IgG1 Fc sequence. In some embodiments, the variant Fc protein, which includes the variant IgG1 Fc protein, includes a cleavage of SEQ ID NO: 26. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the SEQ ID NO: 26. In some embodiments, the cleavage includes the deletion of an amino acid from the N-terminus of SEQ ID NO: 26. In some embodiments, the cleavage includes the deletion of an amino acid from the C-terminus of SEQ ID NO: 26. In some embodiments, the cleavage includes the deletion of an amino acid from both the N-terminus and C-terminus of SEQ ID NO: 26. In some embodiments, the sequence number 26 includes the amino acid sequence of SEQ ID NO: 81: EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(Sequence ID 81)

[0117] In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further comprises one or more mutations selected from the group consisting of L19A, L20A, N82A, P114G, I38A, H95A, and H220A of SEQ ID NO: 81. It should be understood that positions L19, L20, N82, P114, I38, H95, and H220 relate only to SEQ ID NO: 81. Those skilled in the art will readily recognize that positions L234, L235, N297, P329, I253, H310, and H435, numbered according to Kabat's EU numbering system, correspond to positions L19, L20, N82, P114, I38, H95, and H220 of SEQ ID NO: 81, respectively. Any of the mutations L19A, L20A, N82A, P114G, I38A, H95A, and H220A in SEQ ID NO: 81 may or may not be present, and these mutations may be combined in any combination. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further includes mutations corresponding to L19A and L20A of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further includes mutations corresponding to N82A of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further includes mutations corresponding to P114G of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further includes mutations corresponding to L19A, L20A, N82A, and P114G of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further comprises a mutation corresponding to I38A in SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further comprises a mutation corresponding to H95A in SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further comprises a mutation corresponding to H220A in SEQ ID NO: 81.In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further includes mutations corresponding to I38A, H95A, and H220A of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further includes mutations corresponding to L19A, L20A, N82A, P114G, I38A, H95A, and H220A of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 and further including mutations corresponding to L18A, L19A, N82A, P114G, I38A, H95A, and H220A of SEQ ID NO: 81 contains the amino acid sequence of SEQ ID NO: 82. EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMASRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLAQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNAYTQKSLSLSPGK(Sequence ID 82)

[0118] In some embodiments, the variant Fc protein is the variant IgG2 Fc protein (SEQ ID NO: 27). In some embodiments, the variant IgG2 Fc protein contains one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A, numbered according to Kabat's EU numbering index, at the position corresponding to SEQ ID NO: 27. Any of N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 27 may or may not be present, and these mutations may be combined in any combination. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to N297A in SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to P329G in SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutations corresponding to N297A and P329G in SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains a mutation corresponding to I253A in SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains a mutation corresponding to H310A in SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains a mutation corresponding to H435A in SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains mutations corresponding to I253A, H310A, and H435A in SEQ ID NO: 27.

[0119] In some embodiments, the variant Fc protein, which includes the variant IgG2 Fc protein, includes a cleavage of the IgG2 Fc sequence. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the IgG2 Fc sequence. In some embodiments, the variant Fc protein, which includes the variant IgG2 Fc protein, includes a cleavage of SEQ ID NO: 27. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the SEQ ID NO: 27. In some embodiments, the cleavage includes the deletion of an amino acid from the N-terminus of SEQ ID NO: 27. In some embodiments, the cleavage includes the deletion of an amino acid from the C-terminus of SEQ ID NO: 27. In some embodiments, the cleavage includes the deletion of an amino acid from both the N-terminus and C-terminus of SEQ ID NO: 27.

[0120] In some embodiments, the variant Fc protein is the variant IgG4 Fc protein (SEQ ID NO: 28). In some embodiments, the variant IgG4 Fc protein contains one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A, numbered according to Kabat's EU numbering index, at the position corresponding to SEQ ID NO: 28. Any of the mutations S228P, L235E, N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 28 may or may not be present, and these mutations may be combined in any combination. In some embodiments, the variant IgG4 Fc protein contains the mutation corresponding to S228P in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains the mutation corresponding to L235E in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to N297A in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to P329G in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains mutations corresponding to S228P, L235E, N297A, and P329G in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to I253A in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to H310A in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to H435A in SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains mutations corresponding to I253A, H310A, and H435A in SEQ ID NO: 28.

[0121] In some embodiments, the variant Fc protein, which includes the variant IgG4 Fc protein, includes a cleavage of the IgG4 Fc sequence. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the IgG4 Fc sequence. In some embodiments, the variant Fc protein, which includes the variant IgG4 Fc protein, includes a cleavage of SEQ ID NO: 28. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the SEQ ID NO: 28. In some embodiments, the cleavage includes the deletion of an amino acid from the N-terminus of SEQ ID NO: 28. In some embodiments, the cleavage includes the deletion of an amino acid from the C-terminus of SEQ ID NO: 28. In some embodiments, the cleavage includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 28.

[0122] In some embodiments, the stalk moiety S1 comprises a variant Fc protein given by the formula L1-Fc-L2-X1, where L1 is a linker or absent, Fc is a variant Fc protein, L2 is a linker or absent, and X1 is a polypeptide containing a transmembrane domain. Thus, the targeting moiety comprising the formula T-S1 may also be given by the formula T-L1-Fc-L2-X1, where T is a target-binding domain, L1 is a linker or absent, Fc is a variant Fc protein, L2 is a linker or absent, and X1 is a polypeptide containing a transmembrane domain. Therefore, it should be understood that in some embodiments, the stalk moiety S1 may be given by the formula L1-Fc-L2-X1. In some embodiments, the target-binding domain T is as provided herein. In some embodiments, the variant Fc protein is as provided herein.

[0123] In some embodiments, L1 and L2 are polypeptide linkers, each independently. In some embodiments, the polypeptide linker is (GGGGA) n (Sequence No. 54), (GGGGS) n (Sequence ID 55), (EAAAK) n(Sequence ID 73), A(EAAAK) n A (Sequence ID 74), (XP) n (SEQ ID NO: 75) (wherein X is Ala, Lys, or Glu), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESKST (SEQ ID NO: 58), AEAAAKEAAAKA (SEQ ID NO: 76), or combinations thereof, where each n is independently 1 to 5. In some embodiments, each n is independently 1. In some embodiments, each n is independently 2. In some embodiments, each n is independently 3. In some embodiments, each n is independently 4. In some embodiments, each n is independently 5. In some embodiments, each n is independently greater than 5. In some embodiments, L1 is absent. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 1 to 5. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 1. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 2. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 3. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 4. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 5. In some embodiments, L1 is (GGGGA)n (Sequence No. 54) or (GGGGS) n (Sequence ID 55) where each n is independently greater than 5. In some embodiments, L2 is absent. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 1 to 5. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 1. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 2. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 3. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 4. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 5. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n In the formula, each n is independently greater than 5. In some embodiments, L1 is (GGGGA) n (Sequence ID 54), where n is 1. In some embodiments, L1 is (GGGGA) n (Sequence ID 54) where n is 2. In some embodiments, L1 is (GGGGA) n (Sequence ID 54) where n is 3. In some embodiments, L1 is (GGGGA) n(Sequence ID 54), where n is 4. In some embodiments, L1 is (GGGGS) n (Sequence ID 55), where n is 1. In some embodiments, L1 is (GGGGS) n (Sequence ID 55), where n is 2. In some embodiments, L1 is (GGGGS) n (Sequence ID 55), where n is 3. In some embodiments, L1 is (GGGGS) n (Sequence ID 55) where n is 4. In some embodiments, L1 is GSAGSAAGSGEF (Sequence ID 56). In some embodiments, L1 is KESGSVSSEQLAQFRSLD (Sequence ID 57). In some embodiments, L1 is EGKSSGSGSESKST (Sequence ID 58).

[0124] In some embodiments, X1 is ECD-T M - Contains a polypeptide having the formula ICD, wherein ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent. M is the transmembrane domain of a transmembrane protein, and ICD is either an intracellular domain of a protein or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or it is absent. Therefore, in some embodiments, the targeting portion containing formula T-S1, which can also be given by formula T-L1-Fc-L2-X1, is formula T-L1-Fc-L2-ECD-T M -Can also be given by ICD, where T is the target-binding domain, L1 is a linker or is absent, Fc is a variant Fc protein, L2 is a linker or is absent, ECD is the extracellular domain of a cell surface protein or a fragment thereof, or is absent, T M L1-Fc-L2-ECD-T is the transmembrane domain of a transmembrane protein, and ICD is either an intracellular domain of a protein or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or it is absent. Therefore, in some embodiments, the stalk portion S1 is of the formula L1-Fc-L2-ECD-T M-Please understand what can be provided by ICD.

[0125] In some embodiments, the stalk portion S1 does not include a variant Fc region. In some embodiments, the stalk portion S1 is given by the formula L3-X1, where L3 is a mobile peptide linker and X1 is a polypeptide comprising a transmembrane domain provided herein. Thus, in some embodiments, the targeting portion comprising formula T-S1 may also be given by the formula T-L3-X1, where T is a target-binding domain, L3 is a mobile peptide linker, and X1 is a polypeptide comprising a transmembrane domain provided herein. Thus, it should be understood that in some embodiments, the stalk portion S1 may be given by the formula L3-X-1. In some embodiments, the S1 stalk portion is attached to the surface of the virus particle via the transmembrane domain.

[0126] In some embodiments, the mobile peptide linker L3 can be any mobile peptide linker. In some embodiments, L3 is (GGGGA) n (Sequence No. 54), (GGGGS) n(Sequence ID 55), GSAGSAAGSGEF (Sequence ID 56), KESGSVSSEQLAQFRSLD (Sequence ID 57), EGKSSGSGSESKST (Sequence ID 58), or any combination thereof, selected from the group of movable linkers, where each n is an integer independently selected from 1 to 4. In some embodiments, each n is an integer independently selected from 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, or 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, each n is independently greater than 10. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 1. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 2. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 3. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 4. In some embodiments, L3 is (GGGGA) n (Sequence ID 54), where n is 5. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 6. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 7. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 8. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 9. In some embodiments, L3 is (GGGGA) n(SEQ ID NO: 54), and n is 10. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54), and n is greater than 10. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 1. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 2. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 3. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 4. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 5. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 6. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 7. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 8. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 9. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 10. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is greater than 10. In some embodiments, L3 is GSAGSAAGSGEF (SEQ ID NO: 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (SEQ ID NO: 57). In some embodiments, L3 is EGKSSGSGSESKST (SEQ ID NO: 58).

[0127] In some embodiments, the flexible peptide linker L3 can be any flexible peptide linker. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54), (GGGGS) n(SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESKST (SEQ ID NO: 58), or any combination thereof, but not limited thereto, is selected from the group of flexible linkers, wherein each n is independently an integer selected from 1 to 4. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 1. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 2. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 3. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 4. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 1. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 2. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 3. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 4. In some embodiments, L3 is GSAGSAAGSGEF (SEQ ID NO: 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (SEQ ID NO: 57). In some embodiments, L3 is EGKSSGSGSESKST (SEQ ID NO: 58).

[0128] In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54), (GGGGS) n(SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESKST (SEQ ID NO: 58), or any combination thereof, selected from the group of movable linkers, where each n is an integer independently selected from 1, 2, or 4. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 4. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 1. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 2. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 4. In some embodiments, L3 is (GGGGS) n (Sequence ID 55), where n is 1. In some embodiments, L3 is (GGGGS) n (Sequence ID 55), where n is 2. In some embodiments, L3 is (GGGGS) n (Sequence ID 55) where n is 4. In some embodiments, L3 is GSAGSAAGSGEF (Sequence ID 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (Sequence ID 57). In some embodiments, L3 is EGKSSGSGSESKST (Sequence ID 58).

[0129] In some embodiments, X1 is ECD-T M - Contains a polypeptide having the formula ICD, wherein ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent. M is the transmembrane domain of a transmembrane protein, and ICD is either an intracellular domain of a protein or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or it is absent. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which can also be given by formula T-L3-X1, is formula T-L3-ECD-TM -Can also be given by ICD, where T is a target-binding domain, L3 is a mobile peptide linker, ECD is the extracellular domain or fragment thereof of a cell surface protein, or is absent. M ICD is the transmembrane domain of a transmembrane protein, and ICD is either an intracellular domain of a protein or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or it is absent. Therefore, in some embodiments, the stalk portion S1 is of formula L3-ECD-T M -Please understand what can be provided by ICD.

[0130] In some embodiments, the ECD is absent. In some embodiments, the ECD may be any suitable extracellular domain or a fragment thereof. In some embodiments, the ECD may originate from a different protein compared to the transmembrane domain. The ECD domain may be the entire ECD domain or a fragment thereof. In some embodiments, the ECD domain is the CD8 or CD28 ECD domain or a fragment thereof. In some embodiments, the ECD domain is the CD8 ECD domain or a fragment thereof. In some embodiments, the CD8 ECD domain contains the polypeptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the CD8 ECD domain consists of or is essentially the polypeptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain contains a polypeptide that is 25 to 45 amino acids long. In some embodiments, the ECD comprises a polypeptide that is at least or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain is the CD28 ECD domain or a fragment thereof. In some embodiments, the CD28 ECD domain comprises the polypeptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the CD28 ECD domain consists of or is essentially the polypeptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the ECD domain comprises a polypeptide that is 25 to 45 amino acids long.In some embodiments, the ECD comprises a polypeptide that is at least or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60).

[0131] In some embodiments, T M This can be any suitable transmembrane domain or fragment thereof. In some embodiments, T M The domain is CD8 or CD28 T M It is a domain or a fragment thereof. In some embodiments, T M The domain is CD8 T M It is a domain or a fragment thereof. In some embodiments, it is the T of CD8. M The domain contains the polypeptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, the T of CD8 M The domain consists of, or is essentially, the polypeptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M This includes a polypeptide that is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M The domain is CD28 T M It is a domain or a fragment thereof. In some embodiments, CD28 T M The domain contains the polypeptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, CD28 T M The domain consists of or is essentially the polypeptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, T MThis includes a polypeptide that is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62).

[0132] In some embodiments, T M The domain originates from the same protein as ECD. In some embodiments, T M The domain originates from a protein different from ECD. In some embodiments, T M The domain is CD8 or CD28T. M A domain or fragment thereof, and an ECD domain is a CD8 or CD28 ECD domain or fragment thereof. In some embodiments, T M The domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, T MThe domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60).

[0133] In some embodiments, the transmembrane domain is ligated to the intracellular domain (ICD) of a transmembrane protein, or a fragment thereof. In some embodiments, the ICD is absent. In some embodiments, the ICD is T MThe domain originates from the same or a different protein. In some embodiments, the ICD includes an Env integration motif. The Env integration motif is a molecule, such as a polypeptide, that can help facilitate the integration of proteins into the viral envelope. A non-limiting example of an Env integration motif is a polypeptide comprising the amino acid sequence NRVRQGYS (SEQ ID NO: 63). This is a non-limiting example, and other peptide sequences, such as GGTETSQVAPA (SEQ ID NO: 64), may be used. In some embodiments, the Env integration motif includes the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 64, or a combination thereof. In some embodiments, the Env integration motif includes the amino acid sequence of SEQ ID NO: 63. In some embodiments, the Env integration motif includes the amino acid sequence of SEQ ID NO: 64.

[0134] In any embodiment of this specification, the target-binding domain "T" is any polypeptide or polynucleotide that can be used to bind to a desired target. In some embodiments, T is CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3, a glycosylated CD43 epitope expressed in acute leukemia or lymphoma but not in hematopoietic precursors, a glycosylated CD43 epitope expressed in non-hematopoietic cancers, A kinase anchor protein 4 (AKAP-4), adrenoceptor beta 3 (ADRB3), AFP, anaplastic lymphoma kinase Autoantibodies against kinase (ALK), androgen receptor, angiopoietin-binding cell surface receptor 2 (Tie 2), desmoglein 1 (Dsg1), desmoglein 3 (Dsg3), B7H3 (CD276), biotin, bone marrow stromal cell antigen 2 (BST2), BST1 / CD157, cancer / testis antigen 1 (NY-ESO-1), cancer / testis antigen 2 (LAGE-1a), carbonic anhydrase IX (CA1X), carcinoembryonic antigen (CEA), CCCTC binding factor (zinc finger protein)-like (BORIS or Brother of the Regulator of Imprinted Site)Site)), CCR4, CD5, CD19, CD20, CD22, CD24, CD30, CD32 (FCGR2A), CD33, CD34, CD38, CD44v6, CD72, CD79a, CD79b, CD97, CD99, CD123, CD171, CD179a, CD179b-IGLll, CD200R, CD276 / B7H3, CD300 molecule-like family member f (CD300LF), CDH1-CD324, CDH6, CDH17, CDH19, Chromosome X open reading frame 61 (CXORF61), Claudin 6 (CLDN6), Claudin 18.2 (CLD18A2 or CLDN18A.2), CMV pp65, C-MYC epitope tag, Cripto, CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24), CSF2RA (GM-CSFR-alpha), C-type lectin domain family 12 member A (CLEC12A), C-type lectin-like molecule-1 (CLL-1 or CLECL1), cyclin B1, cytochrome P450 IB 1 (CYP1B 1), DLL3, EBV-EBNA3c, EGF-bke module-containing mucin-like hormone receptor-like 2 (EMR2), elongation factor 2 mutated (ELF2M), ephrin B2, ephrin type-A receptor 2 (EphA2), epidermal growth factor receptor EGT receptor (EGFR), Epidermal growth factor receptor variant III (EGFRviii), Epidermal cell adhesion molecule (EPCAM), ERG, ETS translocation variant gene 6 located on chromosome 12p12p (ETV6-AML), IgA receptor Fc fragment (FCAR or CD89), Fc receptor-like 5 (FCRL5), Fibroblast activation protein (FAP), FITC, Fms-like tyrosine kinase 3 (FLT3), Folate receptor alpha (FRa or FR1), Folate receptor beta (FRb), Follicle-stimulating hormone receptor (FSHR), Fos-related antigen 1, Fucosyl-GM1, G protein-coupled receptor class C group 5 member D (GPRC5D), G protein-coupled receptor 20 20, GPR20), GAD, Ganglioside G2 (GD2), Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer), Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(1-4)bDGlcp(1-1)Cer), GD3, GFR alpha 4, Glycoprotein 100 (gp1OO), Glypican-3 (GPC3), Gonadotropin hormone receptor (CGHR or GR), GpA33, GpNMB, GPRC5D, Guanylyl cyclase C (GCC), Heat shock protein 70-2 mutated (mut hsp70-2), Hepatitis A virus cellular receptor 1 (HAVCR1), GloboH glycoceramide hexasaccharide portion (GloboH), High molecular weight-melanoma associated antigenantigen (HMWMAA), HIV1 envelope glycoprotein, HLA, HLA-DOA, HLA-A, HLA-A2, HLA-B, HLA-C, HLA-DM, HLA-DOB, HLA-DP, HLA-DQ, HLA-DR, HLA-G, HTLV1-Tax, Human papillomavirus E6 (HPV E6), Human papillomavirus E7 (HPV E7), Human telomerase reverse transcriptase (hTERT), IgE, IL13Ra2, IL13Ra, Immunoglobulin lambda-like polypeptide 1 (IGLL1), Influenza A hemagglutinin (HA), Insulin-like growth factor 1 receptor (IGF-I) 1) Receptors), interleukin-11 receptor alpha (IL-11Ra), interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2), intestinal carboxylesterase, KIT (CD117), KSHV K8.1, KSHV-gH, LAMP1, regmine, leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), luteinizing hormone receptor (LHR), Lewis (Y) antigen, Lews Ag, Liv 1, K9 locus (LY6K), low-conductance chloride channel, lymphocyte antigen 6 complex, lymphocyte antigen 75 (LY75), lymphocyte-specific protein tyrosine kinase (LCK), mammary gland differentiation antigen (NY-BR-1), T cell-recognized melanoma antigen 1 (MelanA or MARTI (Melanoma antigen recognized by T cells 1)), melanoma-associated antigen 1 (MAGE-A1), melanoma cancer testicular antigen-1 (MAD-CT-1), melanoma cancer testicular antigen-2 (MAD-CT-2), melanoma inhibitor of apoptosis (Melanoma inhibitor of apoptosis (ML-IAP), mesothelin, MPL, cell surface-associated mucin 1 (MUC1), N-acetylglucosaminyl-transferase V (NA17), nectin-4, neural cell adhesion molecule (NCAM), NKG2D, NYBR1, O-acetyl-GD2 ganglioside (OAcGD2), olfactory receptor 51E2 (OR51E2), oncogene fusion protein consisting of breakpoint cluster region (BCR), and Abelson murine leukemia virus oncogene homolog 1.homolog1)(Abl)(bcr-abl), P53 variant, paired box protein Pax-3 (PAX3), paired box protein Pax-5 (PAX5), pannexin 3 (PANX3), PDL1, P-glycoprotein, placenta-specific 1 (PLAC1), platelet-derived growth factor receptor beta (PDGFR (Platelet-derived growth factor receptor)-beta), polysialic acid, proacrosin-binding protein sp32 (OY-TES1), prostase, prostate carcinoma tumor antigen-1 (PCT A-1 (prostate carcinoma tumor antigen-1) or galectin 8), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), prostatic acid phosphatase (Prostatic acid Phosphatase (PAP), prostein, protease serine 21 (testicin or PRSS21), proteasome (prosomal macropine) subunit beta type 9 (LMP2), PTK7, Ras G12V, Ras homolog family member C (RhoC), rat sarcoma (Ras) mutant, advanced glycation end product receptor (RAGE-1), receptor tyrosine kinase-like orphan receptor 1 (ROR1), receptor tyrosine protein kinase ERBB2 or Her-22 / neu, renal ubiquitous 1 (RU1), renal ubiquitous 2 (RU2), sarcoma translocation breakpoint, serine 2 (TMPRSS2) ETS fusion gene, sialyl Lewis adhesion molecule molecule (sLe), SLAMF4, SLAMF6, Slea (CA19.9 or sialyl Lewis antigen), sperm protein 17 (SPA17), squamous cell carcinoma antigen recognized by T cells3) (SART3), Stage-specific embryonic antigen-4 (SSEA-4), STEAP1, Survivin, Synovial sarcoma X breakpoint 2 (SSX2), T cell receptor gamma alternative reading frame protein (TARP), TCR-beta 1 chain, TCR-beta 2 chain, TCR-delta chain, TCR-gamma chain, TCR-gamma-delta, telomerase, TGF-beta R2, Antigen recognized by TNT antibody, Thyroid stimulating hormone receptor (TSHR), Tim1- / HVCR1, Tissue Factor 1 (TF1), Tn ag, Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)), TNF receptor family members B-cell maturation (BCMA), transglutaminase 5 (TGS5), transmembrane protease, TROP2, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related protein (TEM7R), tumor protein p53 (p53), tumor-associated glycoprotein 72 (TAG72), tyrosinase, tyrosinase-related protein 2 (TRP-2), uroplakin 2 (UPK2), vascular endothelial growth factor receptor 2 (VEGFR2), V-myc avian myelocytomatosis viral oncogene (neuroblastoma homolog). (neuroblastoma-derived homolog) (MYCN), Wilms tumor proteinThe target binding domain is any polypeptide, polynucleotide, or fragment thereof that binds to protein (WT1), or X antigen family member 1A (XAGE1). In some embodiments, the target binding domain "T" binds to CD7. In some embodiments, the target binding domain "T" binds to CD8. In some embodiments, the target binding domain "T" is an antibody. In the context of this disclosure, it should be understood that "antibody" refers not only to a "complete" antibody comprising two identical heavy chains, two identical light chains, and two antigen-binding fragments, but also to antibody fragments of any isotype, including but not limited to Fab, Fv, scFv, and Fd fragments, chimeric antibodies, humanized antibodies, single-chain antibodies (scAb), single-domain antibodies (dAb), single-domain heavy-chain antibodies, single-domain light-chain antibodies, bispecific antibodies, multispecific antibodies, and fusion proteins comprising the antigen-binding portion of an antibody and a non-antibody protein. In some embodiments, the antibody is selected from the group including scFv, Fab, VHH, single-domain antibodies, etc. In some embodiments, the antibody is scFv. In some embodiments, the antibody is Fab. In some embodiments, the antibody is VHH. In some embodiments, the antibody is a single-domain antibody.

[0135] In some embodiments, the virus particles provided herein are pseudotyped virus particles. In some embodiments, the virus particles are pseudotyped using viral glycoproteins of Paramyxoviridae viruses. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of morbilliviruses such as measles virus. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of measles virus. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of henipaviruses such as nipah virus, cedar virus, or hendra virus. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of nipah virus. In some embodiments, the polypeptides or antibodies provided herein are linked via a linker to envelope glycoprotein G or H of Paramyxoviridae viruses. In some embodiments, Paramyxoviridae viruses are morbilliviruses such as measles virus. In some embodiments, viruses of the Paramyxoviridae family are henipaviruses such as nipah virus, cedar virus, or hendra virus.

[0136] As provided herein, viruses can be pseudotyped with the VSV-G protein (either its wild-type or a mutant). While not bound by any particular theory, a mutant VSV-G protein containing a mutation at position 182, which can be used to pseudotype a virus (e.g., a lentivirus), can be used to pseudotype a virus and transduce cells when the virus contains a targeting moiety. This mutation inhibits or reduces the VSV-G affinity for its native co-receptor, LDL-R. In some embodiments, the provided mutant VSV-G protein can be used to transduce target cells and deliver heterologous molecules to the target cells.

[0137] In some embodiments, a VSV-G protein is provided with a mutation at position 198 compared to SEQ ID NO: 1, or at position 182 compared to SEQ ID NO: 2. SEQ ID NO: 1 is the full-length protein, and SEQ ID NO: 2 is the ectodomain of the VSV-G protein. The 16-mer signal peptide MKCLLYLAFLFIGVNC (SEQ ID NO: 65), shown at the N-terminus of SEQ ID NO: 1, is cleaved, leaving the protein of SEQ ID NO: 2 intact. Thus, the mutation may be mentioned in the context of SEQ ID NO: 2, but should be understood to also occur in the context of SEQ ID NO: 1, which contains the leader sequence, and therefore would be at a position number 16 more than the position enumerated for SEQ ID NO: 2. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the I182D mutation compared to SEQ ID NO: 2. In some embodiments, the mutation is the I182E mutation compared to SEQ ID NO: 2.

[0138] In some embodiments, a VSV-G protein is provided with a mutation at position 198 compared to SEQ ID NO: 10, or at position 182 compared to SEQ ID NO: 11. SEQ ID NO: 10 is the full-length protein, and SEQ ID NO: 11 is the ectodomain of the VSV-G protein. The 16-mer signal peptide MLSYLIFALVVSPILG (SEQ ID NO: 66) shown at the N-terminus of SEQ ID NO: 10 is cleaved, leaving the protein of SEQ ID NO: 11 intact. Thus, the mutation can be specified for SEQ ID NO: 11, but naturally, it can also be specified for SEQ ID NO: 10. SEQ ID NO: 10 contains this leader sequence and therefore has a position number 16 more than the position shown for SEQ ID NO: 11. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the T182D mutation compared to SEQ ID NO: 11. In some embodiments, the mutation is the T182E mutation compared to SEQ ID NO: 11.

[0139] In some embodiments, a VSV-G protein is provided with a mutation at position 198 compared to SEQ ID NO: 12, or at position 182 compared to SEQ ID NO: 13. SEQ ID NO: 12 is the full-length protein, and SEQ ID NO: 13 is the ectodomain of the VSV-G protein. The 16-mer signal peptide MLRLFLFCFLALGAHS (SEQ ID NO: 67) shown at the N-terminus of SEQ ID NO: 12 is cleaved, leaving the protein of SEQ ID NO: 13 intact. Thus, the mutation can be specified for SEQ ID NO: 13, but naturally, it can also be specified for SEQ ID NO: 12. SEQ ID NO: 12 contains this leader sequence and therefore has a position number 16 more than the position shown for SEQ ID NO: 13. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the A182D mutation compared to SEQ ID NO: 13. In some embodiments, the mutation is the A182E mutation compared to SEQ ID NO: 13.

[0140] In some embodiments, a VSV-G protein is provided with a mutation at position 203 compared to SEQ ID NO: 14, or at position 182 compared to SEQ ID NO: 15. SEQ ID NO: 14 is the full-length protein, and SEQ ID NO: 15 is the ectodomain of the VSV-G protein. The 21-mer signal peptide MKMKMVIAGLILCIGILPAIG (SEQ ID NO: 68), shown at the N-terminus of SEQ ID NO: 14, is cleaved leaving the protein of SEQ ID NO: 15 intact. Thus, the mutation can be specified for SEQ ID NO: 15, but naturally, it can also be specified for SEQ ID NO: 14. SEQ ID NO: 14 contains this leader sequence and therefore has a position number 21 more than the position shown for SEQ ID NO: 15. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the V182D mutation compared to SEQ ID NO: 15. In some embodiments, the mutation is the V182E mutation compared to SEQ ID NO: 15.

[0141] In some embodiments, a VSV-G protein is provided with a mutation at position 199 compared to SEQ ID NO: 16, or at position 182 compared to SEQ ID NO: 17. SEQ ID NO: 16 is the full-length protein, and SEQ ID NO: 17 is the ectodomain of the VSV-G protein. The 17-mer signal peptide MTPAFILCMLLAGSSWA (SEQ ID NO: 69), shown at the N-terminus of SEQ ID NO: 16, is cleaved leaving the protein of SEQ ID NO: 17 intact. Thus, the mutation can be specified for SEQ ID NO: 17, but naturally, it can also be specified for SEQ ID NO: 16. SEQ ID NO: 16 contains this leader sequence and therefore has a position number 17 more than the position shown for SEQ ID NO: 17. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the V182D mutation compared to SEQ ID NO: 17. In some embodiments, the mutation is the V182E mutation compared to SEQ ID NO: 17.

[0142] In some embodiments, a VSV-G protein is provided with a mutation at position 199 compared to SEQ ID NO: 18, or at position 182 compared to SEQ ID NO: 19. SEQ ID NO: 18 is the full-length protein, and SEQ ID NO: 19 is the ectodomain of the VSV-G protein. The 17-mer signal peptide MNFLLLTFIVLPLCSHA (SEQ ID NO: 70), shown at the N-terminus of SEQ ID NO: 18, is cleaved, leaving the protein of SEQ ID NO: 19 intact. Thus, the mutation can be specified for SEQ ID NO: 19, but naturally, it can also be specified for SEQ ID NO: 18. SEQ ID NO: 18 contains this leader sequence and therefore has a position number 17 positions higher than the position shown for SEQ ID NO: 19. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the V182D mutation compared to SEQ ID NO: 19. In some embodiments, the mutation is the V182E mutation compared to SEQ ID NO: 19.

[0143] In some embodiments, a VSV-G protein is provided that contains a mutation at position 199 compared to SEQ ID NO: 20, or at position 182 compared to SEQ ID NO: 21. SEQ ID NO: 20 is the full-length protein, and SEQ ID NO: 21 is the ectodomain of the VSV-G protein. The 17-mer signal peptide MLVLYLLLSLLALGAQC (SEQ ID NO: 71), shown at the N-terminus of SEQ ID NO: 20, is cleaved, leaving the protein of SEQ ID NO: 21 intact. Thus, the mutation can be specified for SEQ ID NO: 21, but naturally, it can also be specified for SEQ ID NO: 20. SEQ ID NO: 20 contains this leader sequence and therefore has a position number 17 positions higher than the position shown for SEQ ID NO: 21. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the I182D mutation compared to SEQ ID NO: 21. In some embodiments, the mutation is the I182E mutation compared to SEQ ID NO: 21.

[0144] Where used herein, when a polypeptide is said to have mutations compared to a reference sequence, such a comparison is based on alignment, such as using BlastP, ClustalW, or ClutalOmega alignment software with default parameters. For example, position 182 can be found in SEQ ID NO: 2 and can also be found in comparison to other strains, as shown in Figure 3. Figure 3 shows cluster alignments of wild-type sequences of the ectodomains of various strains of the VSV-G protein. The residues in bold and underlined are the residues aligned to position 182 of SEQ ID NO: 2 in various strains. SEQ ID NO: 2 refers to the ectodomain of the VSV-G protein from the Indiana strain. SEQ ID NO: 11 refers to the ectodomain of the VSV-G protein from the New Jersey strain. SEQ ID NO: 13 refers to the ectodomain of the VSV-G protein from the Marraba strain. SEQ ID NO: 15 refers to the ectodomain of the VSV-G protein from the Carajas strain. SEQ ID NO: 17 refers to the extracellular domain of the VSV-G protein from the Alagoa strain. Sequence ID 19 refers to the ectodomain of the VSV-G protein of the Cocal strain. Sequence ID 21 refers to the ectodomain of the VSV-G protein of the Morreton strain. Therefore, the residue aligned at residue 182 compared to Sequence ID 2 can also be mutated as provided herein.

[0145] In some embodiments, the mutation at position 182 compared to SEQ ID NO: 2 is not alanine. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 2 is not valine.

[0146] In some embodiments, the mutation at position 182 compared to sequence number 2 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 compared to sequence number 11 is T182S, T182H, T182Q, or T182N. In some embodiments, the mutation at position 182 compared to sequence number 13 is A182S, A182H, A182T, A182Q, or A182N. In some embodiments, the mutation at position 182 compared to sequence number 15 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to sequence number 17 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 19 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 21 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 is not a hydrophobic residue. In some embodiments, the mutation at position 182 is a charged residue. In some embodiments, the mutation at position 182 is a negatively charged residue.

[0147] Mutations may be described with reference to SEQ ID NO: 1 or SEQ ID NO: 2, which are VSV-G proteins derived from the Indiana strain, but mutations can also be used in other strains of the VSV-G protein. For example, mutations can be produced in the VSV-G New Jersey strain, the VSV-G Malaba strain, the VSV-G Carajas strain, the VSV-G Alagoa strain, the VSV-G Kaukal strain, or the VSV-G Moreton strain. In some embodiments, each sequence is provided herein. Examples of these can be found, for example, in U.S. Patent Application Publication No. 20200216502, which is incorporated herein by reference. For example, the wild-type full-length or ectodomain of the VSV-G New Jersey strain is sequence number 10 and sequence number 11, respectively; the wild-type full-length or ectodomain of the VSV-G Marraba strain is sequence number 12 and sequence number 13, respectively; the wild-type full-length or ectodomain of the VSV-G Carajas strain is sequence number 14 and sequence number 15, respectively; the wild-type full-length or ectodomain of the VSV-G Alagoa strain is sequence number 16 and sequence number 17, respectively; the wild-type full-length or ectodomain of the VSV-G Cocal strain is sequence number 18 and sequence number 19, respectively; or the wild-type full-length or ectodomain of the VSV-G Morreton strain is sequence number 20 and sequence number 21, respectively.

[0148] Compared to SEQ ID NO: 2, a VSV-G protein containing a mutation at position 182 may also contain other mutations, such as those described in U.S. Patent Application Publication No. 20200216502 (which is incorporated herein by reference in its entirety). For example, a VSV-G protein may contain mutations at positions corresponding to positions 8, 47, 209, and / or 354 of SEQ ID NO: 2.

[0149] In some embodiments, the substitution at position 8 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for Y. In some embodiments, the substitution at position 209 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for H. In some embodiments, the substitution at position 47 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for K or R. In some embodiments, the substitution at position 354 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for K or R.

[0150] In some embodiments, the substitution is a substitution of A, G, F, or Q at position 47 or 354, or both positions 47 and 354. In some embodiments, the substitution is A or Q.

[0151] In some embodiments, the substitution at position 8 is alanine, i.e., H8A.

[0152] In some embodiments, the substitution at position 47 is Q or N, i.e., K47Q or K47N.

[0153] In some embodiments, the protein contains a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0154] In some embodiments, compared to SEQ ID NO: 2, a protein containing a mutation at position 182 is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 2 (or SEQ ID NO: 1 if using a full-length protein). In some embodiments, the polypeptide contains the I182D or I182E mutation. In some embodiments, the VSV-G protein contains the I182S, I182H, I182T, I182Q, or I182N mutation.

[0155] In some embodiments, a protein containing a mutation at position 182 is identical to SEQ ID NO: 11 by at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% compared to SEQ ID NO: 11 (or SEQ ID NO: 10 if a full-length protein is used). In some embodiments, the polypeptide contains the T182D or T182E mutation. In some embodiments, the VSV-G protein contains the T182S, T182H, T182Q, or T182N mutation.

[0156] In some embodiments, a protein containing a mutation at position 182 is identical to SEQ ID NO: 13 by at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% compared to SEQ ID NO: 13 (or SEQ ID NO: 12 if a full-length protein is used). In some embodiments, the polypeptide contains the A182D or A182E mutation. In some embodiments, the VSV-G protein contains the A182S, A182H, A182T, A182Q, or A182N mutation.

[0157] In some embodiments, a protein containing a mutation at position 182 is identical to SEQ ID NO: 15 by at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% compared to SEQ ID NO: 15 (or SEQ ID NO: 14 if using a full-length protein). In some embodiments, the polypeptide contains the V182D or V182E mutation. In some embodiments, the VSV-G protein contains the V182S, V182H, V182T, V182Q, or V182N mutation.

[0158] In some embodiments, a protein containing a mutation at position 182 is identical to SEQ ID NO: 17 by at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% compared to SEQ ID NO: 17 (or SEQ ID NO: 16 if a full-length protein is used). In some embodiments, the polypeptide contains the V182D or V182E mutation. In some embodiments, the VSV-G protein contains the V182S, V182H, V182T, V182Q, or V182N mutation.

[0159] In some embodiments, a protein containing a mutation at position 182 is identical to SEQ ID NO: 19 by at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% compared to SEQ ID NO: 19 (or SEQ ID NO: 18 if a full-length protein is used). In some embodiments, the polypeptide contains the V182D or V182E mutation. In some embodiments, the VSV-G protein contains the V182S, V182H, V182T, V182Q, or V182N mutation.

[0160] In some embodiments, a protein containing a mutation at position 182 is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 21 (or SEQ ID NO: 20 if a full-length protein is used). In some embodiments, the polypeptide contains the I182D or I182E mutation. In some embodiments, the VSV-G protein contains the I182S, I182H, I182T, I182Q, or I182N mutation.

[0161] Viral glycoproteins The mutant VSV-G protein can be used, for example, to pseudotype viruses such as lentiviruses, without limitation. Accordingly, in some embodiments, viral particles containing the mutant VSV-G protein provided herein are provided. In some embodiments, the viral particles contain a VSV-G protein with a mutation at position 198 compared to SEQ ID NO: 1. In some embodiments, a protein with a mutation at position 182 compared to SEQ ID NO: 2 is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 2 (or SEQ ID NO: 1 if a full-length protein is used). In some embodiments, the polypeptide contains the I182D or I182E mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein contains the I182S, I182H, I182T, I182Q, or I182N mutation.

[0162] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 198 compared to SEQ ID NO: 10. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 11 contains a mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical compared to SEQ ID NO: 11 (or SEQ ID NO: 10 if a full-length protein is used). In some embodiments, the polypeptide contains a T182D or T182E mutation compared to SEQ ID NO: 11. In some embodiments, the VSV-G protein contains T182S, T182H, T182Q, or T182N mutations.

[0163] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 198 compared to SEQ ID NO: 12. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 13 contains a mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical compared to SEQ ID NO: 13 (or SEQ ID NO: 12 if a full-length protein is used). In some embodiments, the polypeptide contains an A182D or A182E mutation compared to SEQ ID NO: 13. In some embodiments, the VSV-G protein includes A182S, A182H, A182T, A182Q, or A182N mutations.

[0164] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 203 compared to SEQ ID NO: 14. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 15 contains a mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical compared to SEQ ID NO: 15 (or SEQ ID NO: 14 if using a full-length protein). In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 15. In some embodiments, the VSV-G protein includes V182S, V182H, V182T, V182Q, or V182N mutations.

[0165] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 199 compared to SEQ ID NO: 16. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 17 contains a mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical compared to SEQ ID NO: 17 (or SEQ ID NO: 16 if a full-length protein is used). In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 17. In some embodiments, the VSV-G protein includes V182S, V182H, V182T, V182Q, or V182N mutations.

[0166] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 199 compared to SEQ ID NO: 18. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 19 contains a mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical compared to SEQ ID NO: 19 (or SEQ ID NO: 18 if a full-length protein is used). In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 19. In some embodiments, the VSV-G protein includes V182S, V182H, V182T, V182Q, or V182N mutations.

[0167] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 199 compared to SEQ ID NO: 20. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 21 contains a mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical compared to SEQ ID NO: 21 (or SEQ ID NO: 20 if a full-length protein is used). In some embodiments, the polypeptide contains an I182D or I182E mutation compared to SEQ ID NO: 21. In some embodiments, the VSV-G protein contains I182S, I182H, I182T, I182Q, or I182N mutations.

[0168] In some embodiments, the VSV-G protein further includes mutations at positions corresponding to position 214 and / or 352 of SEQ ID NO: 2. In some embodiments, the residue corresponding to position 214 of SEQ ID NO: 2 is T214. In some embodiments, the residue corresponding to position 352 of SEQ ID NO: 2 is T352. In some embodiments, the VSV-G polypeptide includes substitutions at position I182 of SEQ ID NO: 2 and at least one of T214 and T352. In some embodiments, the VSV-G polypeptide includes substitutions at positions I182, T214, and T352 of SEQ ID NO: 2. In some embodiments, the VSV-G protein includes a mutation corresponding to the T214N mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein includes a mutation corresponding to the T352A mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein includes the T214N and T352A mutations compared to SEQ ID NO: 2. These mutations can be combined with any other mutations provided herein. In some embodiments, the T214N and / or T352A mutations are combined with the I182E or I182D mutation. In some embodiments, the VSV-G protein contains the amino acid sequences of SEQ ID NOs. 22 and 23, which combine I182D or I182E with the T214N and T352A mutations, respectively. The sequences are also illustrated below along with the leader sequences that are removed during protein processing. VSV-G protein_I196D, T230N, and T368A mutations (with leader sequences and adjusted numbering) MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQS CGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKD LFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 24) VSV-G protein_I182D, T214N, and T352A mutations (no leader sequence) KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAV IVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFP ECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 22) VSV-G protein containing I196E, T230N, and T368A mutations (with leader sequence and adjusted numbering) MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQS CGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKD LFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 25) VSV-G protein containing I182E, T214N, and T352A mutations (no leader sequence) KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAV IVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFP ECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 23)

[0169] In some embodiments, the VSV-G polypeptide includes the sequence of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25. In some embodiments, the VSV-G polypeptide includes the sequence of SEQ ID NO: 22. In some embodiments, the VSV-G polypeptide includes the sequence of SEQ ID NO: 23. In some embodiments, the VSV-G polypeptide includes the sequence of SEQ ID NO: 24. In some embodiments, the VSV-G polypeptide includes the sequence of SEQ ID NO: 25. It should be understood that the VSV-G sequences of SEQ ID NOs. 22-25 are illustrative and not intended to be limiting. Other strains of the VSV-G protein described herein may also include mutations corresponding to T214N and / or T352A in SEQ ID NO: 2, as well as the mutations shown in SEQ ID NOs. 22 and 23. Such additional sequences are within the scope of this disclosure.

[0170] In some embodiments, the composition includes mutations described in Hwang et al., Gene Ther 2013 Aug, 20(8):807-15. (Epub 2013 Jan 31) (which is incorporated herein by reference in its entirety). For example, the mutations may be at positions 230, 368, 66, and / or 162, corresponding to SEQ ID NO: 1. If the leader sequence is removed, the positions are 16 positions fewer compared to SEQ ID NO: 2. In some embodiments, the mutations at these positions are, for example, T230N, T368A, K66T, S162T, or any combination thereof. In some embodiments, the VSV-G protein includes the T230N and T368A mutations. In some embodiments, the VSV-G polypeptide includes K66T, S162T, T230N, and T368A. These positions correspond to positions in the full-length protein (SEQ ID NO: 1). In some embodiments, the VSV-G protein includes the T230N mutation, the T368A mutation, the K66T mutation, the S162T mutation, or any combination thereof. In some embodiments, the VSV-G protein further includes one or more mutations in addition to the mutation corresponding to position 182 of SEQ ID NO: 2, such as that described in U.S. Patent Application Publication No. 20200216502 (which is incorporated herein by reference in its entirety). For example, the VSV-G protein may further include mutations at positions 8, 47, 209, and / or 354 of SEQ ID NO: 2.

[0171] In some embodiments, the substitution at position 8 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for Y. In some embodiments, the substitution at position 209 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for H. In some embodiments, the substitution at position 47 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for K or R. In some embodiments, the substitution at position 354 is by any amino acid different from the amino acid shown at that position in sequence number 2, except for K or R. In some embodiments, the substitution at position 47 or 354, or both positions 47 and 354, is substituted with A, G, F, or Q. In some embodiments, the substitution is A or Q. In some embodiments, the substitution at position 8 is alanine, i.e., H8A. In some embodiments, the substitution at position 47 is Q or N, i.e., K47Q or K47N. In some embodiments, the protein contains a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0172] Furthermore, in some embodiments, the virus can be pseudotyped with other viral structural proteins instead of the VSV-G protein or its variants.

[0173] For example, viral particles can be pseudotyped with Spring viremia of carp virus G (SVCV-G) protein and transduction into cells if the virus contains a targeting moiety. The Spring viremia of carp virus G protein provided can be used in some embodiments to transduce target cells and deliver heterologous molecules to target cells. In some embodiments, the Spring viremia of carp virus G (SVCV-G) protein containing SEQ ID NO: 52 is provided. SEQ ID NO: 52 is a full-length protein, and SEQ ID NO: 53 is the ectodomain of the Spring viremia of carp virus G protein with the N-terminal signal peptide removed. Thus, in some embodiments, the protein contains the amino acid sequence of SEQ ID NO: 53. The Spring viremia of carp virus G protein can be used for pseudotyped viruses, for example, lentiviruses, etc. Thus, in some embodiments, viral particles containing the Spring viremia of carp virus G protein provided herein are provided. In some embodiments, the virus particles contain a carp spring viremia virus G protein comprising a sequence that is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence 52 or 53.

[0174] Sequence of carp spring viremia virus G (sequence number 52 - containing the leader sequence) MSIISYIAFLLLIDSNLGIPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDDLGF PPQSCGWASVTTVSNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWV EKTAGTLTTIHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELD GNIYQGFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0175] Sequence of carp spring viremia virus G (SEQ ID NO: 53 - no leader sequence): IPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDLGFPPQSCGWASVTTV SNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWVEKTAGTLTT IHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELDGNIYQ GFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0176] Gene editing systems In some embodiments, the viral particle includes at least one nucleic acid molecule encoding a gene editing system. The term “gene editing system” refers to some or all of the components required for a functional gene editing complex. For example, the CRISPR / Cas9 gene editing system may include guide RNA or a single guide RNA and the Cas9 protein. In another embodiment, the CRISPR / Cas9 gene editing system may include a single guide RNA and a nucleic acid molecule encoding the Cas9 protein, or multiple molecules. In yet another embodiment, the gene editing system may include a zinc finger nuclease system, a transcription activator-like effector nuclease (TALEN) system, or a meganuclease system.

[0177] In some embodiments, the gene editing system is a CRISPR-Cas system. In some embodiments, the gene editing system is a zinc finger nuclease system. In some embodiments, the gene editing system is a TALEN. In some embodiments, the gene editing system is a meganuclease. In some embodiments, the gene editing system is a gene product regulatory nucleic acid molecule. The gene product that regulates the nucleic acid molecule is known in the art, and any such product may be used. In some embodiments, the gene product regulatory nucleic acid molecule is selected from the group including, but not limited to, siRNA, piRNA, miRNA, RNAi, mRNA, shRNA, and antisense RNA. In some embodiments, the gene product regulatory nucleic acid molecule is siRNA. In some embodiments, the gene product regulatory nucleic acid molecule is piRNA. In some embodiments, the gene product regulatory nucleic acid molecule is miRNA. In some embodiments, the gene product regulatory nucleic acid molecule is RNAi. In some embodiments, the gene product regulatory nucleic acid molecule is mRNA. In some embodiments, the gene product regulatory nucleic acid molecule is shRNA. In some embodiments, the gene product regulatory nucleic acid molecule is antisense RNA.

[0178] In some embodiments, the gene editing system may be a CRISPR-Cas system. The CRISPR-Cas system may include a CRISPR system guide or a polynucleotide encoding the guide. The guide may direct the sequence-specific binding of the CRISPR complex to a target sequence at a genomic locus of the cell being manipulated. The target sequence may be modified by the CRISPR-Cas system. Alternatively or additionally, the CRISPR-Cas system may include a CRISPR-Cas protein or a polynucleotide encoding a CRISPR protein. In some cases, the CRISPR-Cas system may further include a template or a polynucleotide encoding a template. The template may include a nucleic acid sequence that can modify the target sequence. In some embodiments, the nucleic acid molecule encodes a CRISPR-Cas system comprising a Cas protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule. In some embodiments, the CRISPR-Cas system is a Class 1 or Class 2 CRISPR-Cas system. In some embodiments, the CRISPR-Cas system is a Class 1 CRISPR-Cas system. In some embodiments, the CRISPR-Cas system is a Class 2 CRISPR-Cas system. In some embodiments, the Class 2 CRISPR-Cas system includes a Type II Cas protein. In some embodiments, the Type II Cas protein is a Cas9 protein. In some embodiments, the Class 2 CRISPR-Cas system includes a Type V Cas protein. In some embodiments, the Type V Cas protein is selected from the group including, but not limited to, Cas12a, Cas12b, Cas12c, Cas12d, Cas12e, or Cas 14. In some embodiments, the Type V Cas protein is Cas12a. In some embodiments, the Type V Cas protein is Cas12b. In some embodiments, the Type V Cas protein is Cas12c. In some embodiments, the Type V Cas protein is Cas12d. In some embodiments, the Type V Cas protein is Cas12e. In some embodiments, the V-type Cas protein is Cas 14.In some embodiments, the Class 2 CRISPR-Cas system includes a type VI Cas protein. In some embodiments, the type VI Cas protein is selected from the group including, but not limited to, Cas13a, Cas13b, Cas13c, or Cas13d. In some embodiments, the type VI Cas protein is Cas13a. In some embodiments, the type VI Cas protein is Cas13b. In some embodiments, the type VI Cas protein is Cas13c. In some embodiments, the type VI Cas protein is Cas13d. In some embodiments, the CRISPR-Cas system is a base editing system. In some embodiments, the CRISPR-Cas base editing system includes a dCas linked to cytidine deaminase or adenosine deaminase. In some embodiments, the CRISPR-Cas base editing system includes a dCas linked to cytidine deaminase. In some embodiments, the CRISPR-Cas base editing system includes a dCas linked to adenosine deaminase. In some embodiments, the CRISPR-Cas system is a prime editing system. In some embodiments, the CRISPR-Cas prime editing system includes CRISPRa, CRISPRi, Cas 3, or CasMINI protein. In some embodiments, the CRISPR-Cas prime editing system includes CRISPRa. In some embodiments, the CRISPR-Cas prime editing system includes CRISPRi. In some embodiments, the CRISPR-Cas prime editing system includes Cas 3. In some embodiments, the CRISPR-Cas prime editing system includes CasMINI protein.

[0179] In some embodiments, the gene editing system is a TALEN system. For example, a TALEN system may include a transcription activator-like effector nuclease (TALEN), an engineered variant thereof, or a nucleic acid encoding it.

[0180] In some embodiments, the gene editing system is a ZFN system. For example, a ZFN system may include a zinc finger nuclease, an engineered variant thereof, or a nucleic acid encoding it. A ZFN system may use an artificial restriction enzyme produced by fusing a zinc finger DNA-binding domain to a DNA-cleaving domain that can be engineered to target a desired DNA sequence.

[0181] In some embodiments, the gene editing system is a meganuclease system. For example, the meganuclease system may include a meganuclease, an engineered variant thereof, or a nucleic acid encoding it. The meganuclease may be an endodeoxyribonuclease characterized by a large recognition site (a double-stranded DNA sequence of 12-40 base pairs).

[0182] The term “endogenous” is used herein to indicate that a molecule is native to a particular cell, cell population, cell type, tissue, or organism. For example, an endogenous gene refers to a gene that is native to a particular cell, cell population, cell type, tissue, or organism. In some embodiments, the target nucleic acid molecule is edited and is an endogenous gene.

[0183] targeting part In some embodiments, the viral particle comprises a targeting moiety having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. The targeting moiety can be used to target viral particles containing a variant VSV-G protein or SVCV-G protein to cells expressing a target to which the targeting moiety binds. In some embodiments, the target-binding domain is an antibody, scFv antibody, antigen-binding domain, an ankyrin repeat (e.g., DARPIN), VHH domain antibody, nanobody, single-domain antibody, FN3 domain, or any combination thereof. The target-binding domain can be attached to the viral surface via a variant Fc protein (e.g., L1-Fc-L2-X1) as provided herein, or via a mobile polypeptide (e.g., L3-X1) as provided herein. In some embodiments, the identities of L1, L2, L3, Fc, and X1 are as provided herein. In some embodiments, the targeted moiety is attached to (fused to or ligated to) the envelope glycoprotein G or H of a paramyxoviridae virus (e.g., morbillivirus such as measles virus, or henipavirus such as nipah virus, cedarvirus, or hendravirus). In some embodiments, the targeted moiety may be attached to (fused to or ligated to) the glycoprotein of a Rhabdoviridae virus, such as bullous stomatitis New Jersey virus, bullous stomatitis Indiana virus, bullous stomatitis Aragoas virus, bullous stomatitis Maraba virus, bullous stomatitis Karajas virus, parainfluenza virus, Spodoptera frugiperda rhabdovirus isolate Sf G, Drosophila obscura Sigmavirus 10A, Wuhan insect virus 7, perch virus, or koi spring viremia virus. In some embodiments, the VSV protein is a mutant protein such as those provided herein. In some embodiments, the targeting portion is attached to the glycoprotein of a filoviridae virus, such as the Ebola virus, or to the glycoprotein of an arenaviridae virus, such as the Machupo virus.

[0184] In some embodiments, the target-binding domain is scFv. In some embodiments, the target-binding domain is a single-domain antibody. In some embodiments, the target-binding domain is VHH.

[0185] In some embodiments, the targeted portion is CD7, CD8, cKit(CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3, a glycoside expressed in acute leukemia or lymphoma but not in hematopoietic precursors. Autoantibodies against sylated CD43 epitopes, glycosylated CD43 epitopes expressed in non-hematopoietic cancers, A kinase anchor protein 4, adrenergic receptor beta-3 (ADRB3), AFP, anaplastic lymphoma kinase (ALK), androgen receptor, angiopoietin-binding cell surface receptor 2 (Tie2), desmoglein 1 (Dsg1), desmoglein 3 (Dsg3), B7H3 (CD276), biotin, bone marrow stromal cell antigen 2 (BST2) BST1 / CD157, cancer / testicular antigen 1 (NY-ESO-1), cancer / testicular antigen 2 (LAGE-la), carbonic anhydrase IX (CA1X), carcinoembryonic antigen (CEA), CCCTC binding factor (zinc finger protein)-like (BORIS or brother of the regulator of imprinted site), CCR4, CD5, CD19, CD20, CD22, CD24, CD30, CD32 (FCGR2A), CD33, CD34, CD38, CD44v6 CD72, CD79a, CD79b, CD97, CD99, CD123, CD171, CD179a, CD179b-IGLII, CD200R, CD276 / B7H3; CD300 molecule-like family member f (CD300LF), CDH1-CD324, CDH6, CDH17, CDH19, X chromosome open reading frame 61 (CXORF61), claudin 6 (CLDN6), claudin 18.2 (CLD18A2 or CLDN18A.2), CMVpp65, C-MYC epitope tag, Cripto, CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24), CSF2RA (GM-CSFR-alpha), C-type lectin domain family 12 member A (CLEC12A), C-type lectin-like molecule-1 (CLL-1 or CLECL1), cyclin B1, cytochrome P450 IB 1 (CYP1B 1) DLL3, EBV-EBNA3c, EGF-bke module-containing mucin-like hormone receptor 2 (EMR2), elongation factor 2 mutant (ELF2M), ephrin B2, ephrin type A receptor 2 (EphA2), epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRviii), epidermal cell adhesion molecule (EPCAM), ERG, ETS translocation variant gene 6 (ETV6-AML) located on chromosome 12p, IgA receptor Fc fragment (FCAR or CD89), Fc receptor-like 5 (FCRL5), fibroblast-activating protein alpha (FAP), FITC, Fms-like tyrosine kinase 3 (FLT3), folate receptor alpha (FRa or FR1), folate receptor beta (FRb), follicle-stimulating hormone receptor (FSHR) Fos-related antigen 1, fucosyl-GM1, G protein-coupled receptor class C group 5 member D (GPRC5D), G protein-coupled receptor 20 (GPR20), GAD, ganglioside G2 (GD2), ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer), ganglioside GM3 (aNeu5Ac(2-3)bDClalp(1-4)bDGlcp(1-1)Cer), GD3, GFR alpha 4, glycoprotein 100 (gp1OO), glypican-3 (GPC3), gonadotropin hormone receptor (CGHR or GR), GpA33, GpNMB, GPRC5D, guanylyl cyclase C (GCC), heat shock protein 70-2 mutant (mutHSP70-2), Hepatitis A virus cell receptor 1 (HAVCR1), GloboH glycoceramide hexasaccharide portion (GloboH), High molecular weight melanoma-associated antigen (HMWMAA), HIV1 envelope glycoprotein, HLA, HLA-DOA, HLA-A, HLA-A2, HLA-B, HLA-C, HLA-DM, HLA-DOB, HLA-DP, HLA-DQ, HLA-DR, HLA-G, HTLV1-Tax, Human papillomavirus E6 (HPV E6), Human papillomavirus E7 (HPV E7), human telomerase reverse transcriptase (hTERT), IgE, IL13Ra2, IL11Ra, immunoglobulin lambda-like polypeptide 1 (IGLL1), influenza A hemagglutinin (HA), insulin-like growth factor 1 receptor (IGF-I receptor), interleukin-11 receptor alpha (IL-11Ra), interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2), intestinal carboxylesterase, KIT (CD117), KSHV K8.1, KSHV-gH, LAMP1, regmine, leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), luteinizing hormone receptor (LHR), Lewis (Y) antigen, Lewis Ag, Liv1, K9 locus (LY6K), low conductance chloride channel, lymphocyte antigen 6 complex, lymphocyte antigen 75 (LY75), lymphocyte-specific protein tyrosine kinase (LCK), mammary gland differentiation antigen (NY-BR-1), T cell-recognized melanoma antigen 1 (MelanA or MARTI), melanoma-associated antigen 1 (MAGE-A1), melanoma cancer testicular antigen-1 (MAD-CT-1), melanoma cancer testicular antigen-2 (MAD-CT-2), melanoma inhibitor of apoptosisapoptosis (ML-IAP), mesothelin, MPL, cell surface-bound mucin 1 (MUC1), N-acetylglucosaminyl-transferase V (NA17), nectin-4, nerve cell adhesion molecule (NCAM), NKG2D, NYBR1, O-acetyl-GD2 ganglioside (OAcGD2), olfactory receptor 51E2 (OR51E2), oncogene fusion protein consisting of breakpoint cluster region (BCR), and Abelson murine leukemia virus oncogene homolog 1. 1)(Abl)(bcr-abl), P53 variant, paired box protein Pax-3 (PAX3), paired box protein Pax-5 (PAX5), panexin 3 (PANX3), PDL1, P-glycoprotein, placenta-specific 1 (PLAC1), platelet-derived growth factor receptor beta (PDGFR-beta), polysialic acid, proacrosin-binding protein sp32 (OY-TES1), prostase, prostate cancer tumor antigen-1 (PCT A-1 or galectin 8), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), prostatic acid phosphatase (PAP), prostein, protease serine 21 (testicin or PRSS21), proteasome (prosome macropain) subunit beta type 9 (LMP2), PTK7, RasG12V, Ras homolog family member C (RhoC), rat sarcoma (Ras) mutant, advanced glycation end product receptor (RAGE-1), receptor tyrosine kinase-like orphan receptor 1 (ROR1), receptor tyrosine protein kinase ERBB2 or Her-22 / neu, renal ubiquitous receptor 1 (RU1), renal ubiquitous receptor 2 (RU2), sarcoma translocation breakpoint, serine 2 (TMPRSS2) ETS fusion gene, sialyl Lewis adhesion molecule (sLe), SLAMF4, SLAMF6, Slea (CA19.9 or sialyl Lewis antigen), sperm protein 17 (SPA17), squamous cell carcinoma antigen 3 recognized by T cells (SART3), early embryo-specific antigen-4 (SSEA-4), STEAP1, survivor, synovial sarcoma, X-section 2 (SSX2), TCR gamma surrogate leading frame protein (TARP), TCR-beta 1 chain, TCR-beta 2 chain, TCR-delta chain, TCR-gamma chain, TCR-gamma-delta, telomerase, TGF-beta R2, antigen recognized by TNT antibody, thyroid-stimulating hormone receptor (TSHR), Tim1- / HVCR1, tissue factor 1 (TF1), Tn It binds to ag, Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)), TNF receptor family member B cell maturation (BCMA), transglutaminase 5 (TGS5), transmembrane protease, TROP2, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related protein (TEM7R), tumor protein p53 (p53), tumor-associated glycoprotein 72 (TAG72), tyrosinase, tyrosinase-related protein 2 (TRP-2), uroplakin 2 (UPK2), vascular endothelial growth factor receptor 2 (VEGFR2), V-myc avian myelocyte virus oncogene neuroblastoma-derived homolog (MYCN), Wilms tumor protein (WT1), or X antigen family member 1A (XAGE1). In some embodiments, the targeting moiety binds to CD7. In some embodiments, the targeting moiety binds to CD8.

[0186] In some embodiments, the targeting portion binds to a target present on the cell. In some embodiments, the cell is selected from a group that includes, but is not limited to, immune cells, NK cells, dendritic cells, neutrophils, macrophages, or cancer cells. In some embodiments, the immune cells are T cells or B cells. In some embodiments, the targeting portion binds to cells selected from a group including, but not limited to, CD3+ T cells, CD4+ T cells, CD7+ T cells, CD8+ T cells, CD19+ B cells, CD19+ cancer cells, CD20+ B cells, CD20+ cancer cells, CD30+ lung epithelial cells, CD34+ hematopoietic stem cells, CD105+ endothelial cells, CD105+ hematopoietic stem cells, CD117+ hematopoietic stem cells, CD133+ cancer cells, EpCAM+ cancer cells, GluA2+ neurons, GluA4+ neurons, hematopoietic stem cells, hepatocytes, Her2 / Neu+ cancer cells, NKG2D+ natural killer cells, CD7+ NK cells, SLC1A3+ astrocytes, or SLC7A10+ adipocytes. In some embodiments, the cells are T cells. In some embodiments, the cells are B cells. In some embodiments, the cells are CD7+ T cells and / or CD8+ T cells.

[0187] CD7-conjugated polypeptide In some embodiments, the targeting portion (polypeptide) binds to CD7.

[0188] In some embodiments, the polypeptide that binds to CD7 is an antibody that binds to non-human primate CD7. In some embodiments, the polypeptide that binds to CD7 is an antibody that binds to human CD7. The sequence of human CD7 (UniProtKB P09564) is as follows (SEQ ID NO: 29): MAGPPRLLLLPLLLALARGLPGALAAQEVQQSPHCTTVPVGASVNITCSTSGGLRGIYLRQLGPQPQDIIYYEDGVVPTTDRRFRGRIDFSGSQDNLTITMHRLQLSDTGTYTCQAITEVNVYGSGTLVLVTEEQSQGWHRCSDAPPRASALPAPPTGSALPDPQTASALPDPPAASALPAALAVISFLLGLGLGVACVLARTQIKKLCSWRDKNSAACVVYEDMSHSRCNTLSSPNQYQ(Sequence ID 29).

[0189] In some embodiments, the CD7 to which the polypeptide binds is expressed on the surface of the cell. In some embodiments, the cell is an immune cell. In some embodiments, the immune cell is a CD7+ T cell, CD4+ T cell, CD8+ T cell, NK cell, alpha-beta T cell, gamma-delta T cell, lymphoid progenitor cell, hematopoietic stem cell, bone marrow cell, monocyte, macrophage, central memory T cell, effector memory T cell, stem cell-like memory T cell, naive T cell, activated T cell, regulatory T cell (TReg), terminally differentiated effector memory T cell (TEMRA), resident memory T cell (TRM), or T cell CD8+CCR7+.

[0190] In some embodiments, the targeting portion comprises an Fc region and is given by the formula T-L1-Fc-L2-X1. In some embodiments, the targeting portion is an antibody comprising an Fc region. The Fc region may be linked to the heavy or light chain of the antibody. In some embodiments, the Fc region is IgG Fc. In some embodiments, IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, IgG Fc is IgG1 Fc. In some embodiments, the antibody comprises a constant Fc region described herein, such as SEQ ID NOs. 26, 27, or 28, or a variant thereof provided herein. In some embodiments, the identities of T, L1, Fc, L2, and X1 are as provided herein.

[0191] In some embodiments, the targeting portion does not include the Fc region and is given by formula T-L3-X1. In some embodiments, the identities of T, L3, and X1 are as provided herein.

[0192] In some embodiments, polypeptides (e.g., CD7-binding polypeptides) are provided herein. In some embodiments, antibodies (e.g., anti-CD7 antibodies) are provided herein. In some embodiments, the antibody is a recombinant antibody that binds to CD7. In some embodiments, the CD7 protein is a human CD7 protein. In some embodiments, the CD7 protein is a non-human CD7 protein (e.g., mouse, rat, pig, dog, or non-human primate). As used herein, the term “recombinant antibody” refers to an antibody that does not exist in nature. In some embodiments, the term “recombinant antibody” refers to an antibody that has not been isolated from a human subject.

[0193] In some embodiments, an antibody or an antigen-binding fragment thereof is provided, which comprises a peptide selected from the following table, which shows the CDR based on Kabat numbering. TIFF2026518334000002.tif33170

[0194] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to non-human primate CD7. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to human CD7. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having a sequence selected from SEQ ID NOs.33 to 35. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO.33. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO.34. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO.35. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having a sequence selected from SEQ ID NOs.30 to 32. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 30. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 31. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 32. The CDRs referred to throughout the embodiments herein can be replaced with CDRs characterized by different forms such as Chothia and IMGT.

[0195] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain variable region having LCDR1, LCDR2, and LCDR3, wherein LCDR1 has the sequence of SEQ ID NO: 33, LCDR2 has the sequence of SEQ ID NO: 34, and LCDR3 has the sequence of SEQ ID NO: 35.

[0196] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment thereof comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, where HCDR1 has the sequence of SEQ ID NO: 30, HCDR2 has the sequence of SEQ ID NO: 31, and HCDR3 has the sequence of SEQ ID NO: 32.

[0197] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises (i) a light chain having any one of the aforementioned enumerated combinations of LCDR1, LCDR2, and LCDR3 sequences, and (ii) a heavy chain having any one of the aforementioned enumerated combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0198] Different CDR motifs can be combined in any combination, including those not shown in the table above. For example, the following embodiments are provided as non-limiting examples of such combinations.

[0199] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises (i) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 33, the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 34, and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 35; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 30, the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 31, and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 32, or comprises any of the above variants.

[0200] The previous paragraph may refer to CDR under the Kabat family, but equivalent CDR sequences can be used from the naming conventions of IMGT and CHOTHIA.

[0201] In some embodiments, the light chain variable region CDR1 is replaced by any of the other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced by any of the other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced by any of the other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced by any of the other heavy chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced by any of the other heavy chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced by any of the other heavy chain CDR3 sequences.

[0202] In some embodiments, the polypeptide comprises a heavy chain variable region peptide having one of the following sequences or a variant thereof: TIFF2026518334000003.tif28170

[0203] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences or a variant thereof: TIFF2026518334000004.tif24170

[0204] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V of SEQ ID NO: 36 H Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V of SEQ ID NO: 37 L Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 36, or a variant thereof, V L The peptide comprises the sequence of SEQ ID NO: 37, or a variant thereof. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V H Peptides and V L Contains peptides, V HThe peptide contains the sequence of SEQ ID NO: 36, or a variant thereof, V L The peptide comprises the sequence of SEQ ID NO: 37 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to non-human primate CD7. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 36, or a variant thereof, V L The peptide comprises the sequence of SEQ ID NO: 37 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to human CD7. In some embodiments, V H The peptide contains the sequence of SEQ ID NO: 36, V L The peptide contains the sequence of SEQ ID NO: 37.

[0205] V H Array and V L The array is V H Region and V L The region may be linked by a peptide linker in scFv format, but is not limited to any other format. An example of a peptide linker that can be used to link various peptides provided herein is (GGGGS) n Examples include, but are not limited to, (SEQ ID NO: 55) (where each n is independently 1 to 5). In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, the variable region is not linked by a peptide linker. In some embodiments, the polypeptide includes SEQ ID NO: 36 and SEQ ID NO: 37.

[0206] As provided herein, a polypeptide, antibody, or antigen-binding fragment thereof may be a sequence variant.

[0207] The sequence of a polypeptide or antibody can be modified to produce human IgG antibodies. The sequence transformations provided herein can be modified to obtain other types of antibodies. CDRs can also be linked to other antibodies, proteins, or molecules to create antibody fragments that conjugate CD7.

[0208] In some embodiments, the polypeptide or antibody provided herein is a targeting portion on the surface of the manipulated viral particle. In some embodiments, the targeting portion enables binding to target cells. In some embodiments, the targeting portion is a CD7 binding portion, e.g., the polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38. DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS(Sequence ID 38) Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") described in SEQ ID NO: 38 is an antibody or an antigen-binding fragment thereof. In some embodiments, the target-binding domain "T" is an anti-CD7 antibody.

[0209] In some embodiments, the polypeptide or antibody provided herein is a targeting moiety on the surface of an engineered viral particle. In some embodiments, the engineered viral particle is a pseudotyped virus-like particle. In some embodiments, the targeting moiety enables binding to a target cell. In some embodiments, the targeting moiety is a CD7 binding moiety, e.g., a polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 39, QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (Sequence ID 39)

[0210] Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") described in SEQ ID NO: 39 is an antibody, or an antigen-binding fragment thereof. In some embodiments, the target-binding domain "T" is an anti-CD7 antibody. In some embodiments, the anti-CD7 antibody binds to non-human primate CD7. In some embodiments, the anti-CD7 antibody binds to human CD7.

[0211] CD8-bound polypeptide In some embodiments, the targeting portion (polypeptide) can be bound to CD8.

[0212] In some embodiments, the polypeptide binds to CD8. In some embodiments, the polypeptide binds to CD8-alpha. In some embodiments, the polypeptide binds to CD8-beta. In some embodiments, the polypeptide binds to a CD8 heterodimer. In some embodiments, the CD8 heterodimer contains CD8-alpha and CD8-beta subunits. In some embodiments, the polypeptide binds to a CD8-alpha homodimer. In some embodiments, the polypeptide that binds to CD8 is an antibody that binds to non-human primate CD8. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to non-human primate CD8-alpha. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to non-human primate CD8-beta. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to a non-human primate CD8-alpha homodimer. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to a non-human primate CD8 heterodimer. In some embodiments, the polypeptide that binds to CD8 is an antibody that binds to human CD8. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-alpha. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-beta. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-alpha homodimer. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-heterodimer. The sequence of human CD8-alpha (UniProtKB Q8TAW8) is as follows (Sequence ID 40): MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGCYFCSALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV (Sequence ID 40).

[0213] The sequence of human CD8-beta (UniProtKB Q8TD28) is as follows (sequence number 41): MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKISLSNMRIYWLRQRQAPSSDSHHEFLALWDSAKGTIHGEEVEQEKIAVFRDASRFILNLTSVKPEDSGIYFCMIVGSPELTFGKGTQLSVVDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPITLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQLYK (Sequence ID 41).

[0214] In some embodiments, the CD8 to which the polypeptide binds is expressed on the surface of the cell. In some embodiments, the cell is an immune cell. In some embodiments, the immune cell is a CD7+ T cell, CD4+ T cell, CD8+ T cell, NK cell, alpha-beta T cell, gamma-delta T cell, lymphoid progenitor cell, hematopoietic stem cell, bone marrow cell, monocyte, macrophage, central memory T cell, effector memory T cell, stem cell-like memory T cell, naive T cell, activated T cell, regulatory T cell (TReg), terminally differentiated effector memory T cell (TEMRA), resident memory T cell (TRM), or T cell CD8+CCR7+. In some embodiments, the cell is a CD8+ T cell. In some embodiments, the cell is a CD8+ cell.

[0215] In some embodiments, the targeting portion includes an Fc region and is given by the formula T-L1-Fc-L2-X1. In some embodiments, the targeting portion is an antibody containing an Fc region. The Fc region may be linked to the heavy or light chain of the antibody. In some embodiments, the Fc region is IgG Fc. In some embodiments, IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, IgG Fc is IgG1 Fc. In some embodiments, the antibody includes a constant Fc region described herein, such as SEQ ID NOs. 26, 27, or 28, or a variant thereof provided herein. In some embodiments, the identities of T, L1, Fc, L2, and X1 are as provided herein.

[0216] In some embodiments, the targeting portion does not include the Fc region and is given by formula T-L3-X1. In some embodiments, the identities of T, L3, and X1 are as provided herein.

[0217] In some embodiments, polypeptides (e.g., CD8-conjugated polypeptides) are provided herein. In some embodiments, antibodies (e.g., anti-CD8 antibodies) are provided herein. In some embodiments, the antibody is a recombinant antibody that binds to CD8. In some embodiments, the CD8 protein is a human CD8 protein. In some embodiments, the CD8 protein is a non-human CD8 protein (e.g., mouse, rat, pig, dog, or non-human primate). As used herein, the term “recombinant antibody” refers to an antibody that does not exist in nature. In some embodiments, the term “recombinant antibody” refers to an antibody that has not been isolated from a human subject.

[0218] In some embodiments, an antibody or an antigen-binding fragment thereof is provided, which comprises a peptide selected from the following table, which shows the CDR based on Kabat numbering. TIFF2026518334000005.tif33170

[0219] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to non-human primate CD8. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to human CD8. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having a sequence selected from SEQ ID NOs. 45 to 47. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 45. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 46. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 47. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having a sequence selected from SEQ ID NOs. 42 to 44. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 42. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 43. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 44. The CDRs referred to throughout the embodiments herein can be replaced with CDRs characterized by different forms such as Chothia and IMGT.

[0220] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain variable region having LCDR1, LCDR2, and LCDR3, wherein LCDR1 has the sequence of SEQ ID NO: 45, LCDR2 has the sequence of SEQ ID NO: 46, and LCDR3 has the sequence of SEQ ID NO: 47.

[0221] In some embodiments, the polypeptide, antibody, or antibody-bound fragment thereof comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, where HCDR1 has the sequence of SEQ ID NO: 42, HCDR2 has the sequence of SEQ ID NO: 43, and HCDR3 has the sequence of SEQ ID NO: 44.

[0222] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises (i) a light chain having any one of the aforementioned enumerated combinations of LCDR1, LCDR2, and LCDR3 sequences, and (ii) a heavy chain having any one of the aforementioned enumerated combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0223] Different CDR motifs can be combined in any combination, including those not shown in the table above. For example, the following embodiments are provided as non-limiting examples of such combinations.

[0224] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises (i) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 45, the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 46, and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 47; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 42, the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 43, and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 44, or comprises any of the above variants.

[0225] The previous paragraph may refer to CDR under the Kabat family, but equivalent CDR sequences can be used from the naming conventions of IMGT and CHOTHIA.

[0226] In some embodiments, the light chain variable region CDR1 is replaced by any of the other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced by any of the other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced by any of the other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced by any of the other heavy chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced by any of the other heavy chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced by any of the other heavy chain CDR3 sequences.

[0227] In some embodiments, the polypeptide comprises a heavy chain variable region peptide having one of the following sequences or a variant thereof: TIFF2026518334000006.tif28170

[0228] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences or a variant thereof: TIFF2026518334000007.tif24170

[0229] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V of SEQ ID NO: 48 H Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V of SEQ ID NO: 49 L Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 48, or a variant thereof, V L The peptide comprises the sequence of SEQ ID NO: 49, or a variant thereof. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V H Peptides and V L Contains peptides, V HThe peptide contains the sequence of SEQ ID NO: 48, or a variant thereof, V L The peptide comprises the sequence of SEQ ID NO: 49 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to non-human primate CD8. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 48, or a variant thereof, V L The peptide comprises the sequence of SEQ ID NO: 49 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to human CD8. In some embodiments, V H The peptide contains the sequence of SEQ ID NO: 48, V L The peptide contains the sequence of SEQ ID NO: 49.

[0230] V H Array and V L The array is V H Region and V L The region may be linked by a peptide linker in scFv format, but is not limited to any other format. An example of a peptide linker that can be used to link various peptides provided herein is (GGGGS) n Examples include, but are not limited to, (SEQ ID NO: 55) (where each n is independently 1 to 5). In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, the variable region is not linked by a peptide linker. In some embodiments, the polypeptide includes SEQ ID NO: 48 and SEQ ID NO: 49.

[0231] The sequence of a polypeptide or antibody may be modified to produce human IgG antibodies. The sequence transformations provided herein may be modified to produce other types of antibodies. CDRs can also be linked to other antibodies, proteins, or molecules to create antibody fragments that conjugate CD8.

[0232] In some embodiments, the polypeptide or antibody provided herein is a targeting moiety on the surface of the manipulated viral particle. In some embodiments, the targeting moiety enables binding to target cells. In some embodiments, the target-binding domain ("T") is a CD8-binding moiety, e.g., the polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50. NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS(Sequence ID 50) Alternatively, it may be substantially similar to SEQ ID NO: 50, or an active fragment of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") described in SEQ ID NO: 50 is an antibody, or an antigen-binding fragment thereof. In some embodiments, the targeting portion is an anti-CD8 antibody.

[0233] In some embodiments, the polypeptide or antibody provided herein is a targeting moiety on the surface of the manipulated viral particle. In some embodiments, the manipulated viral particle is a pseudotyped virus-like particle. In some embodiments, the targeting moiety enables binding to target cells. In some embodiments, the target-binding domain ("T") is a CD8-binding moiety, e.g., the polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 51. EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (Sequence ID 51)

[0234] Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") described in SEQ ID NO: 51 is an antibody, or an antigen-binding fragment thereof. In some embodiments, the targeting portion is an anti-CD8 antibody. In some embodiments, the anti-CD8 antibody binds to non-human primate CD8. In some embodiments, the anti-CD8 antibody binds to human CD8.

[0235] Targeted portion including the Fc domain In some embodiments, V H and V LThe polypeptide is linked to a stalk portion S1 containing an Fc region. In some embodiments, the Fc region is as provided herein. In some embodiments, the Fc region is a variant Fc region provided herein. Non-restrictive mutations in the Fc region are provided herein. In some embodiments, the variant Fc region includes a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28 provided herein. In some embodiments, the variant of SEQ ID NO: 26 includes one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A provided herein. In some embodiments, the variant of SEQ ID NO: 27 includes one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A provided herein. In some embodiments, the variant of SEQ ID NO: 28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A provided herein. As provided herein, the heavy chain may be linked to the Fc region. In some embodiments, the Fc region further comprises (e.g., linked to) a transmembrane domain. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is or is absent a linker provided herein, Fc is a variant Fc region provided herein, L2 is or is absent a linker provided herein, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M The transmembrane domain provided herein is a transmembrane domain, and the ICD is either an intracellular domain provided herein or absent. Examples of ECDs include, but are not limited to, the CD8 and / or CD28 extracellular domains provided herein. MExamples include, but are not limited to, the CD8 and / or CD28 transmembrane domains provided herein. In some embodiments, X1 is T M It includes, and ECD and ICD are absent. In some embodiments, X1 is ECD, T M It includes, and ICD is not present. In some embodiments, X1 is T M and includes ICD, but does not include ECD. In some embodiments, X1 is ECD, T M , and ICD. It should be understood that in any of the following embodiments, the ECD, ICD, or both may be optionally omitted. Thus, X1 is CD8 and / or CD28 ECD, CD8 and / or CD28 T M Embodiments including an ICD with an Env embedded motif are understood to include the following X1 members: i) CD8 and / or CD28 ECD, CD8 and / or CD28 T M ii) ICDs including the Env built-in motif, CD8 and / or CD28 T M iii) ICDs including an Env built-in motif, and ICDs in which an ECD does not exist, CD8 and / or CD28 ECD, and CD8 and / or CD28 T M and those for which no ICD exists, as well as iv) CD8 and / or CD28 T M and in which neither ECD nor ICD is present. Similarly, X1 is CD8 and / or CD28 T M Embodiments including an ICD that also includes an Env incorporation motif are understood to include the following X1 members: i) CD8 and / or CD28 T M ICDs including Env incorporation motifs, and ii) CD8 and / or CD28 T M And, there is no ICD. Similarly, X1 is CD8 and / or CD28 ECD and CD8 and / or CD28 T M Embodiments including the following X1 members are understood to include: i) CD8 and / or CD28 ECD and CD8 and / or CD28 T Mii) CD8 and / or CD28 T M And, those for which ECD does not exist. Furthermore, the above explanation applies to specific ECDs, T M It should be understood that this also applies to embodiments in which ICD is not described. For example, if X1 is ECD, CD8 and / or CD28 T M Embodiments including ICD are understood to include the following X1 members: ECD, CD8 and / or CD28 T M ii) ICD, and CD8 and / or CD28 T M iii) CD8 and / or CD28 T M , as well as ICDs for which ECD is not present, and iv) CD8 and / or CD28 T M And, ECD and ICD are absent. Similarly, X1 is CD8 and / or CD28 T M Furthermore, embodiments including the ICD are understood to include the following X1 members: i) CD8 and / or CD28 T M and ICD, and ii) CD8 and / or CD28 T M And, those for which ICD is not present. Similarly, X1 is an ECD and CD8 and / or CD28 T M Embodiments including the following X1 members are understood to include: i) ECD and CD8 and / or CD28 T M ii) CD8 and / or CD28 T M and in which ECD does not exist. Unless otherwise explicitly stated, the above embodiments and descriptions are applicable to any of the following embodiments.

[0236] In some embodiments, X1 is CD8 and / or CD28 ECD, T M , and ICD. In some embodiments, X1 is ECD, CD8 and / or CD28 T M , and ICDs. In some embodiments, X1 is a CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and ICD. In some embodiments, X1 is ECD, TM , and ICD, the ICD includes the env embedded motif provided herein. In some embodiments, X1 is CD8 and / or CD28 ECD, T M , and ICD, the ICD includes the env embedded motif provided herein. In some embodiments, X1 is ECD, CD8 and / or CD28 T M , and ICD, the ICD includes the env embedded motif provided herein. In some embodiments, X1 is CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and including ICD, the ICD includes the env embedded motif provided herein.

[0237] In some embodiments, V provided herein H and V L The polypeptide is linked to a stalk portion (S1) containing an Fc region provided herein. In some embodiments, V provided herein H and V L The polypeptide is linked to a stalk portion (S1) which includes an Fc region containing a transmembrane domain provided herein. In some embodiments, the Fc region further containing a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide containing a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is either an intracellular domain provided herein or is absent.

[0238] In some embodiments, V provided herein H and V LPolypeptides are CD8 and / or CD28 ECD, T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD. In some embodiments, V provided herein H and V L Polypeptides are ECD, CD8 and / or CD28 T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD. In some embodiments, V provided herein H and V L Polypeptides are ECD, T M , and an ICD, which is connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, T M , and an ICD, which is connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD that includes an Env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are ECD, CD8 and / or CD28 T M , and an ICD, which is connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD that includes an Env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M, and an ICD, which is connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD that includes an Env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are ECD, T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, and are immobilized on the surface of a virus particle such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, V H and V L Polypeptides bind to immune cells, such as those provided herein.

[0239] In some embodiments, V has the sequence described in Sequence ID No. 36. H V having the peptide and the sequence described in Sequence ID No. 37 L Peptides are ECD, T M , and are connected to a stalk portion (S1) that includes an Fc region (L1-Fc-L2-X1) including the ICD. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 L The peptide is CD8 and / or CD28 ECD, T M , and is connected to a stalk portion (S1) that includes an Fc region (L1-Fc-L2-X1) including the ICD. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 L The peptides are ECD, CD8 and / or CD28 T M , and is connected to a stalk portion (S1) that includes an Fc region (L1-Fc-L2-X1) including the ICD. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 LThe peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and is connected to a stalk portion (S1) that includes an Fc region (L1-Fc-L2-X1) including the ICD. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 L Peptides are ECD, T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 36. H V having the peptide and the sequence described in Sequence ID No. 37 L The peptide is CD8 and / or CD28 ECD, T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 L The peptides are ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in sequence number 36. H V having the peptide and the sequence described in Sequence ID No. 37 L Peptides are ECD, T M, and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, and are immobilized on the surface of a virus particle such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, V H and V L Polypeptides bind to immune cells, such as those provided herein.

[0240] In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptide is ligated to a stalk moiety (S1) containing an Fc region provided herein. In some embodiments, the V contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptide is linked to a stalk moiety (S1) which includes an Fc region containing a transmembrane domain provided herein. In some embodiments, the Fc region containing the transmembrane domain is L -1A polypeptide having the formula -Fc-L2-X1, wherein L1 is or is absent a linker provided herein, Fc is a variant Fc region provided herein, L2 is or is absent a linker provided herein, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide having the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M V is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L Peptides are ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptide is CD8 and / or CD28 ECD, T M, and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptides are ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L Peptides are ECD, T M, and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptide is CD8 and / or CD28 ECD, T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L The peptides are ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 LThe peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 36. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37 L Peptides are ECD, T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, and are immobilized on the surface of a virus particle such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, V H and V L Polypeptides bind to immune cells, such as those provided herein.

[0241] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37.

[0242] In some embodiments, the polypeptide is V H Peptides and VL Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37, provided that V H Peptides and V L The peptide comprises a light chain CDR having the sequence of SEQ ID NOs. 33-35, and / or a heavy chain CDR having the sequence of SEQ ID NOs. 30-32. In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37, provided that V H Peptides and V L The peptide comprises a light chain CDR1 having the sequence of SEQ ID NO: 33, a light chain CDR2 having the sequence of SEQ ID NO: 34, a light chain CDR3 having the sequence of SEQ ID NO: 35, and / or a heavy chain CDR1 having the sequence of SEQ ID NO: 30, a heavy chain CDR2 having the sequence of SEQ ID NO: 31, and a heavy chain CDR3 having the sequence of SEQ ID NO: 32. In some embodiments, V H or V L The CDRs in the chain are as described in the combinations provided herein.

[0243] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V HThe peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37, provided that V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 33, LCDR2 having the sequence of SEQ ID NO: 34, and LCDR3 having the sequence of SEQ ID NO: 35. H The peptides include HCDR1 having the sequence of SEQ ID NO: 30, HCDR2 having the sequence of SEQ ID NO: 31, and HCDR3 having the sequence of SEQ ID NO: 32.

[0244] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37, provided that V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 33 and containing at most one conservative amino acid substitution, LCDR2 having the sequence of SEQ ID NO: 34 and containing at most one conservative amino acid substitution, and LCDR3 having the sequence of SEQ ID NO: 35 and containing at most one conservative amino acid substitution. HThe peptide comprises HCDR1 having the sequence of SEQ ID NO: 30 and containing at most one conservative amino acid substitution, HCDR2 having the sequence of SEQ ID NO: 31 and containing at most one conservative amino acid substitution, and HCDR3 having the sequence of SEQ ID NO: 32 and containing at most one conservative amino acid substitution.

[0245] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 36, V L The peptide contains the sequence of SEQ ID NO: 37.

[0246] In some embodiments, the polypeptides provided herein bind to non-human primate CD7. In some embodiments, the polypeptides provided herein bind to human CD7.

[0247] As provided herein, different polypeptides (V H or V L ) can be linked by a peptide linker, or may not be linked by a peptide linker and instead form a continuous sequence. In some embodiments, the peptide linker is (GGGGS) n The sequence (SEQ ID NO: 55) is included, where each n is independently 1 to 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. The linked peptide form is V H -ZV L or V L -ZV H It can be expressed by the formula, where Z is a peptide linker. In some embodiments, Z is (GGGGS) n(Sequence ID 55) where each n is independently 1 to 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0248] In some embodiments, V L -ZV H A polypeptide comprising a linked peptide represented by the formula includes a heavy chain variable region as described in SEQ ID NO: 36, which is linked to the light chain variable region as described in SEQ ID NO: 37 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via a peptide linker, V H V connected to L The polypeptide containing the following sequence has the sequence shown below: DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS (Sequence ID 38).

[0249] In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 38. In some embodiments, the polypeptide described in SEQ ID NO: 38 is an antibody or its antigen-binding fragment. In some embodiments, the antibody is an anti-CD7 antibody.

[0250] In some embodiments, V H -ZV L A polypeptide comprising a linked peptide represented by the formula includes a light chain variable region as described in SEQ ID NO: 37, which is linked to the heavy chain variable region as described in SEQ ID NO: 36 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via a peptide linker, V L V connected to H The polypeptide containing the following sequence has the sequence shown below: QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (Sequence ID 39).

[0251] In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 39. In some embodiments, the polypeptide described in SEQ ID NO: 39 is an antibody or its antigen-binding fragment. In some embodiments, the antibody is an anti-CD7 antibody. In some embodiments, the anti-CD7 antibody binds to non-human primate CD7. In some embodiments, the anti-CD7 antibody binds to human CD7.

[0252] In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and comprises a stalk portion (S1) that includes an Fc region, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-TM -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes ECD, TM , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and CD8 and / or CD28 ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising an Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes an ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising an Env embedded motif provided herein. In some embodiments, the ICD comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 38, and the ICD, T M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T MThe identity of the ICD is as provided herein. In some embodiments, the polypeptide binds to immune cells such as those provided herein.

[0253] In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and comprises a stalk portion (S1) that includes an Fc region, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes ECD, T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and CD8 and / or CD28 ECD, T M, and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes an ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising an Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, the ICD includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 39, and the ICD, T M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, the polypeptide binds to immune cells such as those provided herein.

[0254] In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 38 and includes a stalk portion (S1) containing an Fc region, such as those provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 38 and includes a stalk portion (S1) containing an Fc region, the Fc region further includes a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further including the transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or absent, and X1 is a polypeptide containing a transmembrane domain provided herein. As provided herein, X1 is of the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 38, ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 38, CD8 and / or CD28 ECD, T M , and also includes a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 38, ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 38, and CD8 and / or CD28 ECD, CD8 and / or CD28 T M, and also includes a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 38, ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in Sequence ID No. 38, CD8 and / or CD28 ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in Sequence ID No. 38, and includes ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in Sequence ID No. 38, and CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) which includes an ICD containing an Env embedded motif provided herein. In some embodiments, the sequence includes the sequence described in sequence number 38, and the ECD, T M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, the polypeptide binds to immune cells such as those provided herein.

[0255] In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 39 and includes a stalk portion (S1) containing an Fc region, such as those provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 39 and includes a stalk portion (S1) containing an Fc region, the Fc region further includes a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further including the transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide containing a transmembrane domain provided herein. As provided herein, X1 is of the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 39, ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, CD8 and / or CD28 ECD, T M , and also includes a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, ECD, CD8 and / or CD28 T M , and also include a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, and CD8 and / or CD28 ECD, CD8 and / or CD28 T M, and also includes a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 39, ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in Sequence ID No. 39, CD8 and / or CD28 ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in Sequence ID No. 39, and includes ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in Sequence ID No. 39, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) which includes an ICD containing an Env embedded motif provided herein.

[0256] In some embodiments, the sequence includes the sequence described in Sequence ID No. 39, ECD, T M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, the polypeptide binds to immune cells such as those provided herein.

[0257] In some embodiments, the polypeptides provided herein comprising the formula T-S1 include a target-binding domain where T is the target-binding domain, S1 is the stalk portion, and comprises an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83. (Sequence No. 83) Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises an amino acid sequence having at least 95% identity with SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises an amino acid sequence having at least 98% identity with SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises the amino acid sequence of SEQ ID NO: 83.

[0258] In some embodiments, a polypeptide provided herein comprising the formula T-S1, wherein T is a target-binding domain and S1 is a stalk portion, comprises an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises the amino acid sequence of SEQ ID NO: 83, the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83.

[0259] In some embodiments, polypeptides provided herein include the formula T-S1, where T is V H and V LThe target-binding domain, which includes S1, is a polypeptide whose stalk portion contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L The stalk portion "S1" of the polypeptide corresponds to amino acids 172-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein comprising formula T-S1 comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L The stalk portion "S1" of the polypeptide corresponds to amino acids 172-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein comprising formula T-S1 comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L The stalk portion "S1" of the polypeptide corresponds to amino acids 172-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein comprising formula T-S1 comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L The amino acids 172-283 of SEQ ID NO: 83 correspond to the amino acids 284-634 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to the amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein comprising formula T-S1 comprises the amino acid sequence of SEQ ID NO: 83, and the target binding domain V HThis corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L This corresponds to amino acids 172-283 of SEQ ID NO: 83, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83.

[0260] In some embodiments, a polypeptide provided herein having the formula T-L1-Fc-L2-X1, where T is a target-binding domain, L1 is a polypeptide linker or absent, Fc is a variant Fc domain provided herein, L2 is a polypeptide linker or absent, and X1 is a polypeptide comprising a transmembrane domain, the polypeptide comprises an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises amino acids 284-301 of SEQ ID NO: 83, Fc comprises amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, the polypeptides provided herein comprising the formula T-L1-Fc-L2-X1 comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises amino acids 284-301 of SEQ ID NO: 83, Fc comprises amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, the polypeptides provided herein comprising the formula T-L1-Fc-L2-X1 comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises amino acids 284-301 of SEQ ID NO: 83, Fc comprises amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83.In some embodiments, the polypeptide provided herein, comprising the formula T-L1-Fc-L2-X1, comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises amino acids 284-301 of SEQ ID NO: 83, Fc comprises amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83.

[0261] In some embodiments, the polypeptides provided herein include the formula T-L1-Fc-L2-X1, where T is V H and V L The target-binding domain includes L1 which is a polypeptide linker or absent, Fc which is a variant Fc domain provided herein, L2 which is a polypeptide linker or absent, and X1 which is a polypeptide containing a transmembrane domain, the polypeptide comprises an amino acid sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, and the target-binding domain is V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. LL1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, the polypeptides provided herein that include the formula T-L1-Fc-L2-X1 contain an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, polypeptides provided herein that include the formula T-L1-Fc-L2-X1 contain an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, polypeptides provided herein that include the formula T-L1-Fc-L2-X1 contain an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, the polypeptide provided herein, comprising the formula T-L1-Fc-L2-X1, comprises the amino acid sequence of SEQ ID NO: 83, and the target-binding domain V HThis corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83.

[0262] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is a target-binding domain, L1 is a polypeptide linker or is absent, Fc is a variant Fc domain provided herein, L2 is a polypeptide linker or is absent, ECD is an extracellular domain, and T M The ICD is a transmembrane domain, and the ICD is an intracellular domain containing the env embedded motif. The polypeptide contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83. The target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, and T M The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ECD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, contain an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, and TM The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, contain an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, and T M The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, contain an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, and T M The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, comprising the formula ICD, comprises the amino acid sequence of SEQ ID NO: 83, the target binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises amino acids 284-301 of SEQ ID NO: 83, Fc comprises amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD comprises amino acids 543-584 of SEQ ID NO: 83, T MThe first component contains amino acids 585-612 of SEQ ID NO: 83, the second component contains amino acids 613-634 of SEQ ID NO: 83, and the third component contains amino acids 627-634 of SEQ ID NO: 83.

[0263] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is V H and V L A target-binding domain comprising, L1 is a polypeptide linker or absent, Fc is a variant Fc domain provided herein, L2 is a polypeptide linker or absent, ECD is an extracellular domain, T M The ICD is a transmembrane domain, and the ICD is an intracellular domain containing the env embedded motif. The polypeptide contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, and the V of the target-binding domain H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, T M The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. LL1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, T M The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, T M The `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, T MThe `ECD` contains amino acids 585-612 of SEQ ID NO: 83, the `ICD` contains amino acids 613-634 of SEQ ID NO: 83, and the `env` embedded motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, which includes the formula ICD, comprises the amino acid sequence of SEQ ID NO: 83, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 83, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains amino acids 284-301 of SEQ ID NO: 83, Fc contains amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 83, T M The first component contains amino acids 585-612 of SEQ ID NO: 83, the second component contains amino acids 613-634 of SEQ ID NO: 83, and the third component contains amino acids 627-634 of SEQ ID NO: 83.

[0264] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is a target-binding domain, L1 is a polypeptide linker or is absent, Fc is a variant Fc domain provided herein, L2 is a polypeptide linker or is absent, ECD is an extracellular domain, and T ML1 is a transmembrane domain, ICD is an intracellular domain containing the env embedded motif, the polypeptide contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, the target binding domain "T" of the polypeptide contains the amino acid sequence of SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc contains the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, including the formula ICD, comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, the target binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc comprises the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M- The polypeptide provided herein, including the formula ICD, comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, the target binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc comprises the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, including the formula ICD, comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, the target binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc comprises the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M- The polypeptide provided herein, including the formula ICD, comprises the amino acid sequence of SEQ ID NO: 83, the target binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 83, L1 comprises the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc comprises the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The sequence contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83.

[0265] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is V H and V L A target-binding domain comprising, L1 is a polypeptide linker or absent, Fc is a variant Fc domain provided herein, L2 is a polypeptide linker or absent, ECD is an extracellular domain, T M The ICD is a transmembrane domain, and the ICD is an intracellular domain containing the env embedded motif. The polypeptide contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 83, and the V of the target-binding domain H This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 83, and the target binding domain V LL1 contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 83; L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83; Fc contains the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83; L2 is absent; ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83; T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 83, and the target binding domain V L L1 contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc contains the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 83, and the target binding domain V HThis contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 83, and the target binding domain V L L1 contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc contains the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, and the target binding domain V H This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 83, and the target binding domain V L L1 contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc contains the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, which includes the formula ICD, comprises the amino acid sequence of SEQ ID NO: 83, and the target binding domain V HThis contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 83, and the target binding domain V L L1 contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 83, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 83, Fc contains the amino acid sequence of SEQ ID NO: 82 and corresponds to amino acids 302-542 of SEQ ID NO: 83, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 83, T M The sequence contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, the ICD contains amino acids 613-634 of SEQ ID NO: 83, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83.

[0266] In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L Peptides are ECD, T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptide is CD8 and / or CD28 ECD, T M , and is coupled to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptides are ECD, CD8 and / or CD28 T M , and is coupled to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 LThe peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and is coupled to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L Peptides are ECD, T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptide is CD8 and / or CD28 ECD, T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptides are ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) that includes an ICD containing an Env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L Peptides are ECD, T M, and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, and are immobilized on the surface of a virus particle such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, V H and V L The peptides bind to immune cells, such as those provided herein.

[0267] In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptide is ligated to a stalk moiety (S1) containing an Fc region provided herein. In some embodiments, the V contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptide is linked to a stalk moiety (S1) which includes an Fc region containing a transmembrane domain provided herein. In some embodiments, the Fc region containing the transmembrane domain is L - The polypeptide has the formula -Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is ECD-TM -May comprise a polypeptide having the formula ICD, wherein ECD is an extracellular domain or fragment thereof provided herein, or is absent. M V is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L Peptides are ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptide is CD8 and / or CD28 ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 LThe peptides are ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L Peptides are ECD, T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing the Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptide is CD8 and / or CD28 ECD, T M, and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptides are ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and an ICD, which is linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD containing an Env embedded motif provided herein. In some embodiments, a V containing a sequence which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 48. H V containing a peptide and a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49 LPeptides are ECD, T M , and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, and are immobilized on the surface of a virus particle such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, V H and V L The peptides bind to immune cells, such as those provided herein.

[0268] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49.

[0269] In some embodiments, V H Peptides and V L A polypeptide containing a peptide is provided, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49. However, V H Peptides and V LThe peptide comprises a light chain CDR having the sequence of SEQ ID NOs. 45-47, and / or a heavy chain CDR having the sequence of SEQ ID NOs. 42-44. In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49, provided that V H Peptides and V L The peptide comprises a light chain CDR1 having the sequence of SEQ ID NO: 45, a light chain CDR2 having the sequence of SEQ ID NO: 46, a light chain CDR3 having the sequence of SEQ ID NO: 47, and / or a heavy chain CDR1 having the sequence of SEQ ID NO: 42, a heavy chain CDR2 having the sequence of SEQ ID NO: 43, and a heavy chain CDR3 having the sequence of SEQ ID NO: 44. In some embodiments, V H or V L The CDRs in the chain are as described in the combinations provided herein.

[0270] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49, provided that V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 45, LCDR2 having the sequence of SEQ ID NO: 46, and LCDR3 having the sequence of SEQ ID NO: 47, VH The peptides include HCDR1 having the sequence of SEQ ID NO: 42, HCDR2 having the sequence of SEQ ID NO: 43, and HCDR3 having the sequence of SEQ ID NO: 44.

[0271] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48, V L The peptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49, provided that V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 45 and containing at most one conservative amino acid substitution, LCDR2 having the sequence of SEQ ID NO: 46 and containing at most one conservative amino acid substitution, and LCDR3 having the sequence of SEQ ID NO: 47 and containing at most one conservative amino acid substitution. H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 42 and containing at most one conservative amino acid substitution, HCDR2 having the sequence of SEQ ID NO: 43 and containing at most one conservative amino acid substitution, and HCDR3 having the sequence of SEQ ID NO: 44 and containing at most one conservative amino acid substitution.

[0272] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 48, V L The peptide contains the sequence of SEQ ID NO: 49.

[0273] In some embodiments, the polypeptides provided herein bind to non-human primate CD8. In some embodiments, the polypeptides provided herein bind to human CD8.

[0274] As provided herein, different polypeptides (V H or V L ) can be linked by a peptide linker, or may not be linked by a peptide linker and instead form a continuous sequence. In some embodiments, the peptide linker is (GGGGS) n The sequence (SEQ ID NO: 55) is included, where each n is independently 1 to 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. The linked peptide form is V H -ZV L or V L -ZV H It can be expressed by the formula, where Z is a peptide linker. In some embodiments, Z is (GGGGS) n (Sequence ID 55) where each n is independently 1 to 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0275] In some embodiments, V L -ZV H A polypeptide comprising a linked peptide represented by the formula includes a heavy chain variable region as described in SEQ ID NO: 48, which is linked to the light chain variable region as described in SEQ ID NO: 49 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, a peptide linker is used to link the V H V connected to L The polypeptide containing the following sequence has the sequence shown below: NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS(Sequence No. 50).

[0276] In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 50. In some embodiments, the polypeptide described in SEQ ID NO: 50 is an antibody or its antigen-binding fragment. In some embodiments, the antibody is an anti-CD8 antibody.

[0277] In some embodiments, V H -ZV L A polypeptide comprising a linked peptide represented by the formula includes a light chain variable region as described in SEQ ID NO: 49, which is linked to the heavy chain variable region as described in SEQ ID NO: 48 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via a peptide linker, V L V connected to H The polypeptide containing the following sequence has the sequence shown below: EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (Sequence ID 51).

[0278] In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 51. In some embodiments, the polypeptide described in SEQ ID NO: 51 is an antibody or its antigen-binding fragment. In some embodiments, the antibody is an anti-CD8 antibody. In some embodiments, the anti-CD8 antibody binds to non-human primate CD8. In some embodiments, the anti-CD8 antibody binds to human CD8.

[0279] In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and comprises a stalk portion (S1) that includes an Fc region, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May comprise a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and includes CD8 and / or CD28 ECD, T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and includes ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and includes ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD containing the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and CD8 and / or CD28 ECD, T M , and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising an Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and includes an ECD, CD8 and / or CD28 T M, and an ICD comprising a stalk portion (S1) comprising an Fc region (L1-Fc-L2-X1) comprising an ICD comprising the Env embedded motif provided herein. In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 50, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and Fc region including ICD L1 The ICD includes a stalk portion (S1) containing -Fc-L2-X1), and the ICD includes an Env embedded motif provided herein. In some embodiments, the ICD includes a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 50, and the ICD includes an Env embedded motif provided herein. M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, such as those provided herein. In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some embodiments, the polypeptide binds to immune cells such as those provided herein.

[0280] In some embodiments, the polypeptide comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 51, and comprises a stalk portion (S1) that includes an Fc region, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a variant Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising...

Claims

1. A viral particle comprising a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system, wherein the targeting moiety is of formula T-S １ The polypeptide comprises having, where T is the target-binding domain and S １ This is the stalk portion, the virus particle.

2. The virus particle according to claim 1, wherein the gene editing system is a CRISPR-Cas system, a zinc finger nuclease system, TALEN, a meganuclease, or a gene product regulatory nucleic acid molecule.

3. The viral particle according to claim 2, wherein the nucleic acid molecule encodes a CRISPR-Cas system comprising a Cas protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, such as a guide RNA, a single guide RNA, or the Cas protein.

4. The virus particle according to claim 2, wherein the CRISPR-Cas system is a Class 1 or Class 2 CRISPR-Cas system.

5. The viral particle according to claim 4, wherein the Class 2 CRISPR-Cas system comprises a type II Cas protein.

6. The virus particle according to claim 5, wherein the type II Cas protein is the Cas9 protein.

7. The viral particle according to claim 4, wherein the Class 2 CRISPR-Cas system comprises a V-type Cas protein.

8. The virus particle according to claim 7, wherein the V-type Cas protein is selected from Cas12a, Cas12b, Cas12c, Cas12d, Cas12e, or Cas14.

9. The viral particle according to claim 4, wherein the Class 2 CRISPR-Cas system comprises a type VI Cas protein.

10. The virus particle according to claim 9, wherein the VI-type Cas protein is selected from Cas13a, Cas13b, Cas13c, or Cas13d.

11. The virus particle according to claim 2, wherein the CRISPR-Cas system is a base editing system.

12. The virus particle according to claim 11, wherein the CRISPR-Cas system comprises a dCas linked to cytidine deaminase or adenosine deaminase.

13. The virus particle according to claim 2, wherein the CRISPR-Cas system is a prime editing system.

14. The virus particle according to claim 2, wherein the CRISPR-Cas system comprises CRISPRRa, CRISPRi, Cas 3, or CasMINI protein.

15. The virus particle according to claim 2, wherein the gene product regulatory nucleic acid molecule encodes at least one molecule selected from siRNA, piRNA, miRNA, RNAiRNA, mRNA, shRNA, and antisense RNA.

16. The aforementioned stalk portion S １ The virus particle according to claim 1, wherein the variant Fc protein comprises a transmembrane domain, the mutant Fc protein comprises at least one effector mutation, and the effector mutation inhibits the interaction between the Fc protein and Fc-interacting proteins such as FcγR, C1q, FcRβ, or FcRn.

17. The aforementioned S １ The virus particle according to claim 16, wherein the stalk portion is attached to the surface of the virus particle via the transmembrane domain.

18. The virus particle according to claim 16, wherein the Fc protein is an IgG1 Fc, IgG2 Fc, or IgG4 Fc protein.

19. The virus particle according to claim 16, wherein the variant Fc protein comprises a variant of the sequence of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO:

28.

20. The virus particle according to claim 16, wherein the variant Fc protein is the variant IgG1 Fc protein.

21. The virus particle according to claim 20, wherein the variant IgG1 Fc protein contains one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A.

22. The virus particle according to claim 20, wherein the variant IgG1 Fc protein comprises an amino acid sequence having at least 80% identity with SEQ ID NO: 82, at least 85% identity with SEQ ID NO: 82, at least 90% identity with SEQ ID NO: 82, at least 95% identity with SEQ ID NO: 82, at least 98% identity with SEQ ID NO: 82, or at least 100% identity with SEQ ID NO:

82.

23. the formula T-S １ where the targeting moiety having １ L-Fc-L ２ -X １ has a stalk moiety S of the formula １ wherein L １ However, it is either a linker or does not exist. Fc is a variant Fc protein, L ２ However, it is either a linker or does not exist. X １ However, it is a polypeptide that contains a transmembrane domain, The above formula T-S １ The targeting portion having T-L １ -Fc-L ２ -X １ A virus particle according to claim 1, having the formula:

24. L １ and L ２ The virus particle according to claim 23, wherein each of them is either absent independently or is a polypeptide linker containing the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, or SEQ ID NO:

76.

25. X １ However, ECD-T Ｍ - Containing a polypeptide having the formula ICD, in the formula, ECD is either the extracellular domain or fragment thereof of a cell surface protein, or it is absent. T Ｍ However, it is the transmembrane domain of a transmembrane protein. ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, or it is not present. Said T-L １ -Fc-L ２ -X １ The targeting portion having the formula T-L １ -Fc-L ２ -ECD-T Ｍ - A virus particle according to claim 24, having the formula ICD.

26. The virus particle according to claim 25, wherein the ECD comprises the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO:

60.

27. Said T Ｍ The virus particle according to claim 25, wherein the virus particle comprises the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO:

62.

28. The virus particle according to claim 25, wherein the ICD includes an Env embedded motif comprising the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO:

64.

29. The target-binding domain (T) may be CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3, a glycosylated CD43 epitope expressed in acute leukemia or lymphoma but not in hematopoietic precursors, a glycosylated CD43 epitope expressed in non-hematopoietic cancers, A kinase anchor protein 4 (AKAP-4), adrenergic receptor beta-3 (ADRB3), AFP, anaplastic lymphoma kinase (ALK), androgen receptor, angiopoietin-binding cell surface receptor 2 (Tie). 2) Autoantibodies against desmoglein 1 (Dsg1), autoantibodies against desmoglein 3 (Dsg3), B7H3 (CD276), biotin, bone marrow stromal cell antigen 2 (BST2), BST1 / CD157, cancer / testis antigen 1 (NY-ESO-1), cancer / testis antigen 2 (LAGE-1a), carbonic anhydrase IX (CA1X), carcinoembryonic antigen (CEA), CCCTC binding factor (zinc finger protein)-like (BORIS or brother of the regulator of imprinted site), CCR4, CD5, CD19, CD20, CD22, CD24 CD30, CD32 (FCGR2A), CD33, CD34, CD38, CD44v6, CD72, CD79a, CD79b, CD97, CD99, CD123, CD171, CD179a, CD179b-IGLL, CD200R, CD276 / B7H3, CD300 molecule-like family member f (CD300LF), CDH1-CD324, CDH6, CDH17, CDH19, X chromosome open reading frame 61 (CXORF61), claudin 6 (CLDN6), claudin 18.2 (CLD18A2 or CLDN18A.2), CMVpp65, C-MYC epitope tag, crypto, CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24), CSF2RA (GM-CSFR-alpha), C-type lectin domain family 12 member A (CLEC12A), C-type lectin-like molecule-1 (CLL-1 or CLECL1), cyclin B1, cytochrome P450 IB 1 (CYP1B 1) DLL3, EBV-EBNA3c, EGF-bke module-containing mucin-like hormone receptor 2 (EMR2), elongation factor 2 mutant (ELF2M), ephrin B2, ephrin type A receptor 2 (EphA2), epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRviiii), epidermal cell adhesion molecule (EPCAM), ERG, ETS translocation variant gene 6 located on chromosome 12p (ETV6-AML), IgA receptor Fc fragment (FCAR or CD89), Fc receptor-like 5 (FCRL5), fibroblast-activating protein alpha (FAP), FITC, Fms-like tyrosine kinase 3 (FLT3), folate receptor alpha (FRa or FR1), folate receptor beta (FRb), follicle-stimulating hormone receptor (FSH R), Fos-related antigen 1, fucosyl-GM1, G protein-coupled receptor class C group 5 member D (GPRC5D), G protein-coupled receptor 20 (GPR20), GAD, ganglioside G2 (GD2), ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer), ganglioside GM3 (aNeu5Ac(2-3)bDCLarp(1-4)bDGlcp(1-1)Cer), GD3, GFR alpha 4, glycoprotein 100 (gp1OO), glypican-3 (GPC3), gonadotropin hormone receptor (CGHR or GR), GpA33, GpNMB, GPRC5D, guanylyl cyclase C (GCC), heat shock protein 70-2 variant (mutHSP70-2), Hepatitis A virus cell receptor 1 (HAVCR1), GloboH glycoceramide hexasaccharide portion (GloboH), High molecular weight melanoma-associated antigen (HMWMAA), HIV1 envelope glycoprotein, HLA, HLA-DOA, HLA-A, HLA-A2, HLA-B, HLA-C, HLA-DM, HLA-DOB, HLA-DP, HLA-DQ, HLA-DR, HLA-G, HTLV1-Tax, Human papillomavirus E6 (HPV E6), Human papillomavirus E7 (HPV E7), human telomerase reverse transcriptase (hTERT), IgE, IL13Ra2, IL11Ra, immunoglobulin lambda-like polypeptide 1 (IGLL1), influenza A hemagglutinin (HA), insulin-like growth factor 1 receptor (IGF-I receptor), interleukin-11 receptor alpha (IL-11Ra), interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2), intestinal carboxylesterase, KIT (CD117), KSHV K8.1, KSHV-gH, LAMP1, regmine, leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), luteinizing hormone receptor (LHR), Lewis (Y) antigen, Lewis Ag, Liv1, K9 locus (LY6K), low-conductance chloride channel, lymphocyte antigen 6 complex, lymphocyte antigen 75 (LY75), lymphocyte-specific protein tyrosine kinase (LCK), mammary gland differentiation antigen (NY-BR-1), T cell-recognizing melanoma antigen 1 (MelanA or MARTI), melanoma-associated antigen 1 (MAGE-A1), melanoma cancer testicular antigen-1 (MAD-CT-1), melanoma cancer testicular antigen-2 (MAD-CT-2), melanoma inhibitor of apoptosis (ML-IAP), mesothelin, MPL, cell surface-bound mucin 1 (MUC1), N-acetylglucosaminyl-transferaseV (NA17), nectin-4, nerve cell adhesion molecule (NCAM), NKG2D, NYBR1, O-acetyl-GD2 ganglioside (OAcGD2), olfactory receptor 51E2 (OR51E2), oncogene fusion protein (BCR) consisting of cleavage cluster regions, and Abelson mouse leukemia virus oncogene homolog 1 (Abl) (bcr-abl), P53 variant, paired box protein Pax-3 (PAX3), paired box protein Pax-5 (PAX5), panexin 3 (PANX3), PDL1, P-glycoprotein, placenta-specific 1 (PLAC1), platelet-derived growth factor receptor beta (PDGFR-beta), polysialic acid, proacrosin-binding protein sp32 (OY-TES1), prostase, prostate cancer tumor antigen-1 (PCT A-1 or galectin 8), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), prostatic acid phosphatase (PAP), prostein, protease serine 21 (testicin or PRSS21), proteasome (prosomal macropain) subunit beta type 9 (LMP2), PTK7, Ras G12V, Ras homolog family member C (RhoC), rat sarcoma (Ras) mutant, advanced glycation end product receptor (RAGE-1), receptor tyrosine kinase-like orphan receptor 1 (ROR1), receptor tyrosine protein kinase ERBB2 or Her-22 / neu, renal ubiquitous receptor 1 (RU1), renal ubiquitous receptor 2 (RU2), sarcoma translocation breakpoint, serine 2 (TMPRSS2) ETS fusion gene, sialyl Lewis adhesion molecule (sLe), SLAMF4, SLAMF6, Slea (CA19.9 or sialyl Lewis antigen), sperm protein 17 (SPA17), squamous cell carcinoma antigen 3 recognized by T cells (SART3), early embryo-specific antigen-4 (SSEA-4), STEAP1, Survivin, synovial sarcoma X-section 2 (SSX2), TCR gamma surrogate leading frame protein (TARP), TCR-beta 1 chain, TCR-beta 2 chain, TCR-delta chain, TCR-gamma chain, TCR-gamma-delta, telomerase, TGF-beta R2, antigen recognized by TNT antibody, thyroid-stimulating hormone receptor (TSHR), Tim1- / HVCR1, tissue factor 1 (TF1), Tn ag, Tn antigen ((TnA viral particle according to claim 1, which binds to Ag) or (GalNAca-Ser / Thr), TNF receptor family member B cell maturation (BCMA), transglutaminase 5 (TGS5), transmembrane protease, TROP2, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related (TEM7R), tumor protein p53 (p53), tumor-associated glycoprotein 72 (TAG72), tyrosinase, tyrosinase-related protein 2 (TRP-2), uroplakin 2 (UPK2), vascular endothelial growth factor receptor 2 (VEGFR2), V-myc avian myelocyte virus oncogene neuroblastoma-derived homolog (MYCN), Wilms tumor protein (WT1), or X antigen family member 1A (XAGE1).

30. The virus particle according to claim 29, wherein the target-binding domain (T) binds to CD7.

31. The virus particle according to claim 30, wherein the polypeptide comprises a heavy chain and a light chain, the heavy chain having a heavy chain variable region having at least 90% sequence identity with SEQ ID NO: 36, and a light chain having at least 90% sequence identity with SEQ ID NO: 37, and the polypeptide comprises the sequences of HCDR1 described in SEQ ID NO: 30, HCDR2 described in SEQ ID NO: 31, HCDR3 described in SEQ ID NO: 32, LCDR1 described in SEQ ID NO: 33, LCDR2 described in SEQ ID NO: 34, and LCDR3 described in SEQ ID NO:

35.

32. The virus particle according to claim 30, wherein the target binding domain (T) that binds to CD7 comprises a sequence having at least 90% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, at least 95% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, at least 99% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, or a polypeptide comprising the sequence described in SEQ ID NO: 38 or SEQ ID NO:

39.

33. The virus particle according to claim 1, wherein the heterologous viral glycoprotein is a VSV-G polypeptide.

34. The virus particle according to claim 33, wherein the VSV-G polypeptide comprises the sequence of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO:

25.

35. The virus particle according to claim 1, further comprising a nucleic acid molecule encoding a heterogeneous molecule of interest, wherein the heterogeneous molecule of interest is siRNA, shRNA, non-coding RNA (e.g., guide RNA for the CRISPR system), peptide, polypeptide, protein, viral payload, viral genome, or a combination thereof.

36. A method for infecting cells, comprising bringing the cells into contact with a virus particle described in any one of claims 1 to 35.

37. A method for infecting cells in a target, comprising administering to the target a pharmaceutical composition containing virus particles according to any one of claims 1 to 35.

38. A method for editing a target nucleic acid molecule within a cell, comprising contacting the cell with a virus particle described in any one of claims 1 to 35, wherein the nucleic acid molecule encoding the gene editing system is expressed within the cell and edits the target nucleic acid molecule within the cell.

39. The method according to claim 38, wherein the nucleic acid molecule encoding the gene editing system encodes a CRISPR / CAS gene editing system comprising a CAS nuclease and a gRNA that targets the CAS nuclease to the target nucleic acid molecule.

40. The method according to claim 38 or 39, wherein the target nucleic acid molecule to be edited is an endogenous gene.

41. The method according to any one of claims 36 to 40, wherein the cells are T cells, B cells, NK cells, dendritic cells, neutrophils, macrophages, or cancer cells.

42. A method for treating a disease or disorder in a subject requiring treatment of the disease or disorder, comprising administering a viral particle according to any one of claims 1 to 35 to the subject, wherein the nucleic acid molecule encoding the gene editing system is expressed in the subject, and the subject is treated for the disease or disorder.

43. A method for editing a target nucleic acid molecule within a cell, comprising contacting the cell with a viral particle, wherein the viral particle (a) A VSV-G polypeptide having an amino acid sequence that is at least 70% identical to SEQ ID NO: 2, and having a mutation at position 182 compared to SEQ ID NO: 2, (b) A nucleic acid molecule encoding a gene editing system, wherein the gene editing system is a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, (c) By arbitrary selection, a nucleic acid molecule encoding the target heterologous molecule and (d) comprising a targeting portion that binds to T cells, CD3+ T cells, CD4+ T cells, CD7+ T cells, or CD8+ T cells, A method comprising expressing the nucleic acid molecule encoding the gene editing system within the cell and editing the target nucleic acid molecule within the cell.

44. The method according to claim 43, wherein the cells are T cells, CD3+ T cells, CD4+ T cells, CD7+ T cells, CD8+ T cells, or CD7+ NK cells.

45. A viral particle comprising a heterologous viral glycoprotein, a targeting region, and at least one nucleic acid molecule encoding a gene editing system, The heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO:

53. The targeting moiety has the formula T-S １ The polypeptide comprises having, where T is the target-binding domain and S １ This is the stalk part, The target-binding domain includes a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO:

51. The aforementioned stalk portion S １ However, it contains a variant Fc protein that includes a sequence which is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO:

28. The variant of Sequence ID No. 26 includes one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. The variant of Sequence ID No. 27 includes one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A. The variant of Sequence ID No. 28 includes one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A, and The variant Fc protein further comprises a transmembrane domain containing a sequence selected from SEQ ID NO: 61 or SEQ ID NO: 62, and A viral particle in which the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or the Cas protein.

46. A viral particle comprising a heterologous viral glycoprotein, a targeting region, and at least one nucleic acid molecule encoding a gene editing system, The heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO:

53. The targeting moiety has the formula T-S １ The polypeptide comprises having, where T is the target-binding domain and S １ This is the stalk part, The target-binding domain includes a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO:

51. The aforementioned stalk portion S １ However, L １ -Fc-L ２ -X １ The formula includes, in the formula L １ However, the linker either contains an array selected from sequence numbers 54, 55, 56, 57, 58, 73, 74, 75, or 76, or does not exist. Fc is a variant Fc protein containing a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO:

28. The variant of Sequence ID No. 26 includes one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. The variant of Sequence ID No. 27 includes one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A. The variant of Sequence ID No. 28 includes one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. L ２ However, the linker either contains an array selected from sequence numbers 54, 55, 56, 57, 58, 73, 74, 75, or 76, or does not exist. X １ However, ECD-T Ｍ - A polypeptide containing a transmembrane domain having the formula ICD. The ECD is an extracellular domain having a sequence selected from SEQ ID NO: 59 or SEQ ID NO: 60 or a fragment thereof, or it is not present. T Ｍ However, it is a transmembrane domain having the sequence of SEQ ID NO: 61 or SEQ ID NO: 62, or a fragment thereof, and The ICD is an intracellular domain or protein that facilitates the incorporation of the targeted portion into the envelope of the virus particle, and the ICD contains an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent, and A viral particle in which the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or the Cas protein.

47. A viral particle comprising a heterologous viral glycoprotein, a targeting region, and at least one nucleic acid molecule encoding a gene editing system, The heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO:

53. The targeting moiety has the formula T-S １ The polypeptide comprises having, where T is the target-binding domain and S １ This is the stalk part, The target-binding domain includes a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO:

51. The aforementioned stalk portion S １ However, L １ -Fc-L ２ -X １ The formula includes, in the formula L １ However, the linker either contains the sequence of sequence number 55 or does not exist. Fc is a variant Fc protein containing a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO:

28. The variant of Sequence ID No. 26 includes one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. The variant of Sequence ID No. 27 includes one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A. The variant of Sequence ID No. 28 includes one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. L ２ However, the linker either contains the sequence of sequence number 55 or does not exist, and X １ However, ECD-T Ｍ - A polypeptide containing a transmembrane domain having the formula ICD. ECD is either an extracellular domain having the sequence of sequence number 60, or a fragment thereof, or is absent. T Ｍ However, it is a transmembrane domain having the sequence of sequence number 62, The ICD is an intracellular domain or protein that facilitates the incorporation of the targeted portion into the envelope of the virus particle, and the ICD contains an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent, and A viral particle in which the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or the Cas protein.

48. A viral particle comprising a heterologous viral glycoprotein, a targeting region, and at least one nucleic acid molecule encoding a gene editing system, The aforementioned heterologous viral glycoprotein includes the sequence of SEQ ID NO: 23 or SEQ ID NO: 25, The targeting moiety has the formula T-S １ The polypeptide comprises having, where T is the target-binding domain and S １ This is the stalk part, The target-binding domain includes the sequence of Sequence ID No. 39, The aforementioned stalk portion S １ However, L １ -Fc-L ２ -X １ The formula includes, in the formula L １ However, it is a linker that includes the array of sequence number 55, Fc is a variant Fc protein containing the sequence of SEQ ID NO:

82. L ２ However, it is a linker and does not exist. X １ However, ECD-T Ｍ - A polypeptide containing a transmembrane domain having the formula ICD, ECD is an extracellular domain having the sequence of Sequence ID No. 60, T Ｍ However, it is a transmembrane domain having the sequence of sequence number 62, The ICD is an intracellular domain or protein that facilitates the incorporation of the targeted portion into the envelope of the virus particle, and the ICD includes an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63, and A viral particle in which the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or the Cas protein.

49. A viral particle comprising a heterologous viral glycoprotein, a targeting region, and at least one nucleic acid molecule encoding a gene editing system, The heterologous viral glycoprotein contains an amino acid sequence that has at least 90% identity with SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identity with SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identity with SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identity with SEQ ID NO: 23 or SEQ ID NO:

25. The targeted portion includes an amino acid sequence having at least 90% identity with SEQ ID NO: 83, at least 95% identity with SEQ ID NO: 83, at least 99% identity with SEQ ID NO: 83, or at least 100% identity with SEQ ID NO: 83, and A viral particle in which the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule, or a guide RNA, a single guide RNA, or the Cas protein.

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