Gene editing composition of polypeptide and nucleic acid and application thereof

Abstract

Provided herein are viral particles comprising a heterologous viral structural protein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system, compositions comprising these viral particles, and methods of using these viral particles.

Description

[0001] Related applications

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 504,068, filed May 24, 2023, which is incorporated herein by reference in its entirety.

[0003] Reference sequence list submitted electronically

[0004] This application includes a sequence list, which is submitted electronically in XML format and incorporated herein by reference in its entirety. The XML copy created on May 8, 2024, is named “INH-022WO_SL” and has a size of 95,139 bytes. Technical Field

[0005] The implementation schemes provided in this article relate to compositions of peptides and nucleic acids capable of gene editing and methods of using them. Background Technology

[0006] Vesicular stomatitis virus (VSV) is an enveloped, negative-sense RNA virus belonging to the genus Vesiculovirus of the family Rhabdovirus. This virus is an arbovirus that can infect insects, cattle, horses, and pigs. The VSV genome encodes five structural proteins, among which is a single transmembrane glycoprotein (G). Glycoproteins are classic type I membrane glycoproteins with an N-terminal signal peptide, an extracellular domain of approximately 450 amino acids, a single α-helix transmembrane segment, and a small intraviral C-terminal domain. The signal peptide is cleaved in the lumen of the endoplasmic reticulum, and the native glycoprotein is present in the extracellular domain, transmembrane domain, and intraviral domain.

[0007] G plays a key role during the initial stages of virus infection (Albertini, A. A. V., Baquero, E., Ferlin, A., and Gaudin, Y. (2012). Molecular and Cellular Aspects of Rhabdovirus Entry. Viruses 4, 117-139.), which is hereby incorporated by reference in its entirety. First, this glycoprotein is responsible for the attachment of the virus to specific receptors. After binding, the virion enters the cell through a clathrin-mediated endocytosis pathway. In the acidic environment of the endocytic vesicle, G triggers the fusion between the viral and endosomal membranes, which releases the genome in the cytosol for subsequent infection steps. Fusion is catalyzed by a large structural transition from a pre-fusion to a post-fusion conformation, both being trimers, induced by low pH (Roche, S., Bressanelli, S., Rey, F. A., 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, F. A., Gaudin, Y., and Bressanelli, S. (2007). Structure of the prefusion form of the vesicular stomatitis virus glycoprotein g. Science 315, 843-848), each of which is hereby incorporated by reference in its entirety).

[0008] 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, the FD, PHD, and TrD maintain their tertiary structure. However, these domains undergo a large rearrangement in their relative orientation due to a secondary change of the refolding hinge segment (S1 to S5) during the low-pH-induced conformational change (Roche et al., 2006; Roche et al., 2007).

[0009] It has been shown that the low-density lipoprotein receptor (LDL-R) and other members of this receptor family act 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 hereby incorporated 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 tropism as VSV. However, this broad tropism can inhibit selective targeting of specific cell types. Thus, there is a need for modified (mutated or mutant) VSV-G proteins that eliminate their binding to LDL receptors that can be used to pseudotype viruses. Embodiments of the present invention satisfy these needs and others. SUMMARY

[0010] In some embodiments, provided herein is a viral particle. 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 comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety.

[0011] In some embodiments, the stalk moiety 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, wherein the effector mutation inhibits interaction between the Fc protein and an Fc interacting protein, such as FcyR, Clq, FcRb, or FcRn.

[0012] In some embodiments, the variant Fc protein is a variant IgGl Fc protein comprising one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A.

[0013] In some embodiments, the variant IgGl Fc protein comprises an amino acid sequence that is at least 80% identical to SEQ ID NO: 82, at least 85% identical to SEQ ID NO: 82, at least 90% identical to SEQ ID NO: 82, at least 95% identical to SEQ ID NO: 82, at least 98% identical to SEQ ID NO: 82, or at least 100% identical to SEQ ID NO: 82.

[0014] In some embodiments, the variant Fc protein is a variant IgG2 Fc protein comprising one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A.

[0015] In some embodiments, the variant Fc protein is a variant IgG4 Fc protein comprising one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A.

[0016] In some embodiments, the targeting moiety having the formula T-S1 comprises a stalk moiety S1 having the formula L1-Fc-L2-X1, wherein L1 is a linker or is absent, Fc is a variant Fc protein, L2 is a linker or is absent, and X1 is a polypeptide comprising a transmembrane domain.

[0017] In some embodiments, the polypeptide comprising a transmembrane domain (X1) comprises a polypeptide having the formula ECD-T M -ICD, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M is a transmembrane domain of a transmembrane protein; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of a viral particle, or is absent; wherein the targeting moiety having the formula T-L1-Fc-L2-X1 has the formula T-L1-Fc-L2-ECD-T M -ICD.

[0018] In some embodiments, the stalk moiety S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker and X1 is a polypeptide comprising a transmembrane domain; wherein the targeting moiety having the formula T-S1 has the formula T-L3-X1.

[0019] In some embodiments, the polypeptide comprising a transmembrane domain (X1) comprises a polypeptide having the formula ECD-T M -ICD, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T Mis a transmembrane domain of a transmembrane protein; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of a viral particle, or is absent; wherein the targeting moiety having the formula T-L3-X1 has the formula T-L3-ECD-T M - ICD.

[0020] In some embodiments, the targeting moiety binds to CD7. In some embodiments, the targeting moiety 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 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 a variant of any of the foregoing; 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 a variant of any of the foregoing as provided herein.

[0021] In some embodiments, the targeting moiety binds to CD8. In some embodiments, the targeting moiety comprises a polypeptide comprising: 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 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 a variant of any of the foregoing; 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 a variant of any of the foregoing as provided herein.

[0022] In some embodiments, the heterologous viral glycoprotein is a SVCV-G polypeptide. In some embodiments, the SVCV-G polypeptide is as provided herein.

[0023] 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 comprises a substitution at positions I182, T214, and T352 of SEQ ID NO: 2. In some embodiments, the substitution at position 182 is I182D or I182E. In some embodiments, the substitution at position 214 is T214N. In some embodiments, the substitution at position 352 is T352A.

[0024] 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 modulating nucleic acid molecule. 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 a Cas protein to a target nucleic acid molecule. In some embodiments, the gene product modulating nucleic acid molecule encodes at least one molecule selected from the group consisting of an siRNA, a piRNA, a miRNA, an RNAiRNA, a mRNA, a shRNA, and an antisense RNA.

[0025] In some embodiments, a method of infecting a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle as provided herein.

[0026] In some embodiments, a method of infecting a cell of a subject is provided. In some embodiments, the method comprises administering to the subject a pharmaceutical composition comprising a viral particle as provided herein.

[0027] In some embodiments, a method of editing a target nucleic acid molecule in a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle as provided herein, wherein a nucleic acid molecule encoding a gene editing system is expressed in the cell and a target nucleic acid molecule is edited in the cell.

[0028] In some embodiments, a method of treating a disease or condition in a subject is provided. In some embodiments, the method comprises administering to the subject a viral particle as provided herein, wherein a nucleic acid molecule encoding a gene editing system is expressed in the subject and a disease or condition in the subject is treated.

[0029] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises (a) a VSV-G polypeptide comprising an amino acid sequence that is at least 70% identical to SEQ ID NO: 2 and comprises 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 a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets 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, a CD3+ T cell, a CD4+ T cell, a CD7+ T cell, or a CD8+ T cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0030] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises (a) a VSV-G polypeptide comprising an amino acid sequence that is at least 70% identical to SEQ ID NO: 2 and comprises 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 a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets 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, a CD3+ T cell, a CD4+ T cell, a CD7+ T cell, or a CD8+ T cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0031] In some embodiments, a method of editing a target nucleic acid in a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle, wherein the viral particle comprises (a) a VSV-G polypeptide comprising an amino acid sequence that is at least 70% identical to SEQ ID NO: 2 and 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 a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets 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, a CD3+ T cell, a CD4+ T cell, a CD7+ T cell, or a CD8+ T cell; wherein the nucleic acid molecule encoding the gene editing system is expressed in the cell and edits the target nucleic acid molecule in the cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0032] In embodiments, a method of editing a target nucleic acid in a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle, wherein the viral particle comprises (a) a SVCV-G polypeptide comprising a polypeptide 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%, 99%, or 100% identical to SEQ ID NO: 52 or SEQ ID NO: 53; (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 a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets 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, a CD3+ T cell, a CD4+ T cell, a CD7+ T cell, or a CD8+ T cell; wherein the nucleic acid molecule encoding the gene editing system is expressed in the cell and edits the target nucleic acid molecule in the cell. In some embodiments, the heterologous molecule of interest is a chimeric antigen receptor.

[0033] 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 the group consisting of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, and 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 moiety. In some embodiments, the target binding domain comprises an 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 comprising a sequence of SEQ ID NO: 61 or SEQ ID NO: 62. In some embodiments, the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets a Cas protein to a target nucleic acid molecule.

[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 a sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, and 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 moiety. In some embodiments, the target binding domain comprises an 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 L1-Fc-L2-X1, wherein L1 is a linker comprising a sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID O: 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, 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, wherein 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, wherein the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A, wherein 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 a sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID O: 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 X1 is a polypeptide having the formula ECD-T M- a polypeptide of the ICD comprising a transmembrane domain. In some embodiments, the 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, the T M 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 a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, wherein the ICD comprises an env incorporation motif comprising 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 a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets a Cas protein to a target nucleic acid molecule.

[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 selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, and 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 moiety. In some embodiments, the target binding domain comprises an 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 L1-Fc-L2-X1, wherein L1 is a linker comprising a 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, wherein 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, wherein the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A, wherein 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 a sequence of SEQ ID NO: 55, or is absent, and X1 is a polypeptide having the formula ECD-T M -ICD comprising a transmembrane domain. In some embodiments, the ECD is an extracellular domain having the sequence of SEQ ID NO: 60, or is a fragment thereof, or is absent. In some embodiments, T M is a transmembrane domain having the sequence of SEQ ID NO: 62, or is a fragment thereof. In some embodiments, the ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, wherein the ICD comprises an env incorporation motif comprising an 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 a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets a Cas protein to a target nucleic acid molecule.

[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 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, wherein 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, wherein 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 a polypeptide having the formula ECD-T M -ICD comprising a transmembrane domain. In some embodiments, the ECD is an extracellular domain having the sequence of SEQ ID NO: 60. In some embodiments, T M is a transmembrane domain having the sequence of SEQ ID NO: 62. In some embodiments, the ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, wherein the ICD comprises an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63. In some embodiments, the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets a Cas protein to a target nucleic acid molecule.

[0037] 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 that is at least 90% identical to SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identical to SEQ ID NO: 23 or SEQ ID NO: 25, at least 98% identical to SEQ ID NO: 23 or SEQ ID NO: 25, or at least 90% identical to SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the targeting moiety comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 83, at least 95% identical to SEQ ID NO: 83, at least 98% identical to SEQ ID NO: 83, or at least 100% identical to SEQ ID NO: 83. In some embodiments, the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets a Cas protein to a target nucleic acid molecule.

[0038] 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 that is at least 90% identical to SEQ ID NO: 52 or SEQ ID NO: 53, at least 95% identical to SEQ ID NO: 52 or SEQ ID NO: 53, at least 98% identical to SEQ ID NO: 52 or SEQ ID NO: 53, or at least 90% identical to SEQ ID NO: 52 or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 83, at least 95% identical to SEQ ID NO: 83, at least 98% identical to SEQ ID NO: 83, or at least 100% identical to SEQ ID NO: 83. In some embodiments, the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets a Cas protein to a target nucleic acid molecule.

[0039] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting portion, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the targeting portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion. 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 stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible 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 linker having the formula ECD-T. M -ICD is a polypeptide containing a transmembrane domain. In some embodiments, the ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or may not be present. In some embodiments, T M It 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 a protein that facilitates the incorporation of the target portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or is absent. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising the Cas9 protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule.

[0040] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting portion, and at least one nucleic acid molecule encoding a gene editing system. In some embodiments, the targeting portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion. 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 stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible 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 linker having the formula ECD-T. M-ICD is a polypeptide containing a transmembrane domain. In some embodiments, the ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or may not be present. In some embodiments, T M It 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 a protein that facilitates the incorporation of the target portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or is absent. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising the Cas9 protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule.

[0041] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting portion, 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 portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the target-binding domain comprises an 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 stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible linker comprising an 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 polypeptide having the formula ECD-T. M -ICD is a polypeptide containing a transmembrane domain. In some embodiments, the ECD is an extracellular domain or a fragment thereof containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or it is absent. In some embodiments, T MIt is a transmembrane domain or fragment thereof 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 a protein that facilitates the incorporation of the targeting portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or is absent. In some embodiments, the nucleic acid molecule encoding the gene editing system encodes a CRISPR-Cas system comprising the Cas9 protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule.

[0042] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting portion, 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 portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the target-binding domain comprises an 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 stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a linker having the formula ECD-T. M -ICD is a polypeptide containing a transmembrane domain. In some embodiments, ECD is an extracellular domain or a fragment thereof containing the amino acid sequence of SEQ ID NO: 59. In some embodiments, T M It is a transmembrane domain or fragment thereof containing the amino acid sequence of SEQ ID NO: 61. In some embodiments, the ICD is an intracellular domain or a protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, wherein the ICD contains 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 the Cas9 protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule.

[0043] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting portion, 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 portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a linker having the formula ECD-T. M -ICD is a polypeptide containing a transmembrane domain. In some embodiments, ECD is an extracellular domain or a fragment thereof containing the amino acid sequence of SEQ ID NO: 59. In some embodiments, T M It is a transmembrane domain or fragment thereof containing the amino acid sequence of SEQ ID NO: 61. In some embodiments, the ICD is an intracellular domain or a protein that facilitates the incorporation of a targeting portion into the viral particle envelope, wherein the ICD contains 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 the Cas9 protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule.

[0044] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral structural protein, a targeting portion, 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 portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a linker having the formula ECD-T. M -ICD is a polypeptide containing a transmembrane domain. In some embodiments, ECD is an extracellular domain or a fragment thereof containing the amino acid sequence of SEQ ID NO: 59. In some embodiments, T MIt is a transmembrane domain or fragment thereof containing the amino acid sequence of SEQ ID NO: 61. In some embodiments, the ICD is an intracellular domain or a protein that facilitates the incorporation of a targeting portion into the viral particle envelope, wherein the ICD contains 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 the Cas9 protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to the target nucleic acid molecule. Attached Figure Description

[0045] Figure 1A and Figure 1B The crystal structure of VSV-G combined with LDL-R is illustrated. Figure 1A The crystal structure of VSV-G bound to CR3 of LDL-R is illustrated. Figure 1B The crystal structure of VSV-G with CR2 bonded to LDL-R is illustrated.

[0046] Figure 2A and Figure 2B The effect of adding negatively charged amino acids to the VSV-G:LDL-R binding interface on native affinity and cohesion is illustrated. Figure 2A The titration of the VSV-G construct on SupT1 cells is illustrated. Figure 2B Examples are given by Figure 2A The functional titer of each construct is calculated by titration.

[0047] Figure 3 The comparison of extracellular domains of different VSV-G proteins from different strains is illustrated.

[0048] Figure 4 The effects of various VSV-G mutations on the serum stability of viral constructs are illustrated.

[0049] Figure 5 The effects of various VSV-G mutations on the serum stability of viral constructs combined with CD7 binding compounds are illustrated.

[0050] Figure 6 The ability of various rhabdoviral G proteins, alone or in combination with CD7 conjugates, to transduce SupT1 and PBMC cells is illustrated.

[0051] Figures 7A-7L Flow cytometry data of human PBMCs transduced with an exemplary vector containing a CD7 conjugate as disclosed herein are shown.

[0052] Figure 7MFlow cytometry data of human PBMCs transduced with an exemplary vector containing a CD7 conjugate as disclosed herein are shown.

[0053] Figures 8A-8L Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing CD7 conjugates as disclosed herein are shown.

[0054] Figure 8M Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing CD7 conjugates as disclosed herein are shown.

[0055] Figure 9A Flow cytometry data of human PBMCs transduced with an exemplary vector containing a CD8 conjugate as disclosed herein are shown.

[0056] Figure 9B Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing CD8 conjugates as disclosed herein are shown.

[0057] Figure 10A The ability of lentiviral particles carrying CD7 conjugates with mutant Fc stems, pseudotyped VSV-G*, to transduce SupT1 cells and human and non-human primate PBMCs is illustrated. Figure 10B The transduction of cells in the absence of CD7 binders is illustrated. Figure 10C Transduction of human and non-human primate PBMCs is illustrated based on MOI calculated by SupT1 titration. VSV-G* indicates VSV-G (I182E, T214N, T352A).

[0058] Figure 11A The ability of lentiviral particles pseudotyped with SVCV-G carrying CD7 conjugates with mutant Fc stems to transduce SupT1 cells and human and non-human primate PBMCs is illustrated. Figure 11B The transduction of cells in the absence of CD7 binders is illustrated. Figure 11C Transduction of human and non-human primate PBMCs based on MOI calculated by SupT1 titration is illustrated.

[0059] Figures 12A-12D This is a comparison of the transduction capabilities of VSV-G* pseudotyped lentiviral particles (Figures A and B) and SVCV-G pseudotyped lentiviral particles (Figures C and D) into SupT1 cells and human and non-human primate PBMCs. VSV-G* represents VSV-G (I182E, T214N, T352A).

[0060] Figure 13A and Figure 13BThis is a comparison of off-target transduction of GFP in a group of B cell lines compared to the control SupT1 cells. Figure 13A Data for lentiviral particles pseudotyped as VSV-G* carrying CD7 conjugates with mutant Fc stems are illustrated. VSV-G* represents VSV-G (I182E, T214N, T352A). Figure 13B Data are illustrated for lentiviral particles pseudotyped with SVCV-G carrying CD7 conjugates with mutant Fc stems.

[0061] Figure 14A and Figure 14B This is a comparison of off-target transduction of the CAR20-T2A-GFP construct in a group of B cell lines compared to the control SupT1 cells. Figure 14A Data for lentiviral particles pseudotyped as VSV-G* carrying CD7 conjugates with mutant Fc stems are illustrated. VSV-G* represents VSV-G (I182E, T214N, T352A). Figure 14B Data are illustrated for lentiviral particles pseudotyped with SVCV-G carrying CD7 conjugates with mutant Fc stems.

[0062] Figure 15A The ability of lentiviral particles carrying CD7 conjugates with flexible stems, pseudotyped VSV-G*, to transduce SupT1 cells and human and non-human primate PBMCs is illustrated. Figure 15B The transduction of cells in the absence of CD7 binders is illustrated. Figure 15C Transduction of human and non-human primate PBMCs is illustrated based on MOI calculated by SupT1 titration. VSV-G* indicates VSV-G (I182E, T214N, T352A).

[0063] Figure 16A This demonstrates the ability of CD7 conjugates carrying flexible stems of varying lengths to transduce SupT1 cells, compared to other IgG-based conjugates. Figure 16B This demonstrates the ability of CD7 conjugates carrying flexible stems of varying lengths to transduce activated PBMCs compared to other IgG-based conjugates.

[0064] Figure 17A The ability of lentiviral particles carrying CD7 conjugates with flexible stems, pseudotyped SVCV-G, to transduce SupT1 cells and human and non-human primate PBMCs is illustrated. Figure 17B The transduction of cells in the absence of CD7 binders is illustrated. Figure 17C Transduction of human and non-human primate PBMCs based on MOI calculated by SupT1 titration is illustrated.

[0065] Figures 18A-18D This is a comparison of the transduction capabilities of VSV-G* pseudotyped lentiviral particles (Figures A and B) and SVCV-G pseudotyped lentiviral particles (Figures C and D) into SupT1 cells and human and non-human primate PBMCs. VSV-G* represents VSV-G (I182E, T214N, T352A).

[0066] Figure 19A and Figure 19B This is a comparison of off-target transduction of GFP in a group of B cell lines compared to the control SupT1. Figure 19A Data for lentiviral particles pseudotyped as VSV-G* carrying CD7 conjugates with flexible stems are illustrated. VSV-G* represents VSV-G (I182E, T214N, T352A). Figure 19B Data on lentiviral particles carrying CD7 conjugates with flexible stems and pseudotyped SVCV-G are illustrated.

[0067] Figure 20A and Figure 20B This is a comparison of off-target transduction of the CAR20-T2A-GFP construct in a set of B cell lines compared to the control SupT1. Figure 20A Data for lentiviral particles pseudotyped as VSV-G* carrying CD7 conjugates with flexible stems are illustrated. VSV-G* represents VSV-G (I182E, T214N, T352A). Figure 20B Data on lentiviral particles carrying CD7 conjugates with flexible stems and pseudotyped SVCV-G are illustrated.

[0068] Figure 21A and Figure 21B This illustrates the reduction in the percentage of cells positive for the target gene when transduced using the lentiviral constructs of this disclosure. Figure 21A This illustrates the reduction in GFP-positive cells when treated with a lentiviral construct carrying Cas9 and GFP sgRNA. No reduction in GFP-positive cells was observed when the lentiviral construct delivered CD7 sgRNA. Figure 21B This illustrates the reduction in CD7-positive cells when treated with a lentiviral construct carrying either Cas9 or one of two independent CD7 sgRNAs. No reduction in CD7-positive cells was observed when the lentiviral construct delivered GFP sgRNA. Data are based on treatment with 100 µL of virus 7 days post-transduction.

[0069] Figures 22A-22L The presence of different GFP-negative or CD7-negative cell populations at 19 days post-transduction is illustrated. Figures 22A-22D The comparison was made using 100 µL of virus containing CD7 sgRNA (Figure 22A and Figure 22B ) or viruses containing GFP sgRNA ( Figure 22C Samples treated with GFP and untreated parental GFP-positive and CD7-positive cells ( Figure 22D ). Figures 22E-22H The comparison was made using 20 µL of virus containing CD7 sgRNA ( Figure 22E and Figure 22F ) or viruses containing GFP sgRNA ( Figure 22G Samples treated with GFP and untreated parental GFP-positive and CD7-positive cells ( Figure 22H ). Figures 22I-22L The comparison was made using 4 µL of virus containing CD7sgRNA ( Figure 22I and Figure 22J ) or viruses containing GFP sgRNA ( Figure 22K Samples treated with GFP and untreated parental GFP-positive and CD7-positive cells ( Figure 22L ).

[0070] Figure 23 GFP expression in cells treated with either a virus containing CD7 sgRNA or a virus containing GFP sgRNA, compared to parental GFP-positive and CD7-positive cells, is illustrated. Neither the CD7 sgRNA construct resulted in a reduction in GFP expression. However, the GFP sgRNA construct delivered with the viral construct of this disclosure resulted in a robust and observable reduction in GFP expression. Wells marked with an asterisk indicate the harvesting of these cells for further expansion.

[0071] Figure 24 The results of sequence analysis of genomic DNA around the expected Cas9 cleavage site in cells transduced with GFP sgRNA are illustrated. The analysis shows that the GFP gene is cleaved at the expected cleavage site indicated by the guide sequence.

[0072] Figure 25 The results of sequence analysis of genomic DNA around the expected Cas9 cleavage site in cells transduced with CD7_3 sgRNA are illustrated. The analysis shows that the CD7 gene is cleaved at the expected cleavage site indicated by the guide sequence. Detailed Implementation

[0073] This article provides viral particles that may, for example, contain a heterologous viral glycoprotein, a targeting moiety, and at least one nucleic acid molecule encoding a gene editing system. The targeting moiety may be present in the form of T-L1-Fc-L2-X1 or T-L3-X1, where: T is the targeting moiety; L1 is a linker or is absent; Fc is a mutant Fc protein; L2 is a linker or is absent; L3 is a flexible peptide linker; and X1 is a polypeptide containing a transmembrane domain. The mutant Fc polypeptide may be incorporated into the viral particles to help facilitate the targeting of the viral particles to specific cell types. At least one nucleic acid molecule encoding a gene editing system may encode a gene editing system, such as a CRISPR-Cas system containing a Cas protein and guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule.

[0074] Additionally, viral particles may contain VSV-G proteins that can be used, for example, to pseudotype viruses such as lentiviruses. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of vesicular stomatitis New Jersey virus strain, vesicular stomatitis Indiana virus strain, vesicular stomatitis Aragos virus strain, vesicular stomatitis Malaba virus strain, or vesicular stomatitis Carajás virus strain. Examples of such proteins are provided herein.

[0075] Pseudogenized viruses containing mutant VSV-G proteins (such as those provided herein) can be used in conjunction with targeting moieties to promote the fusion of the pseudotyped virus with specific cells or tissues based on the expression of the target on the cell or tissue. As provided herein, the targeting moieties can be linked to an Fc protein, which may refer to a stem protein containing a transmembrane domain to facilitate attachment of the targeting moieties to the viral surface. In some embodiments, the Fc protein contains an Fc effector mutation, such as those provided herein. As provided herein, the targeting moieties can alternatively be linked to a flexible polypeptide, which may be referred to as a “flexible stem protein” or “flexible stem” containing a transmembrane domain to facilitate attachment of the targeting moieties to the viral surface. In some embodiments, the flexible polypeptide is a flexible peptide linker, such as those provided herein.

[0076] Unless otherwise defined, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed embodiments pertain.

[0077] As used herein, unless the context clearly indicates otherwise, the term "a" means "at least one" or "one or more".

[0078] As used herein, the term “about” means an approximate index value, and small variations will not significantly affect the practice of the disclosed embodiments. When numerical limits are used, the “about” index value may vary by ±10% and remain within the range of the disclosed embodiments unless the context otherwise specifies. Furthermore, when the phrase “about x to y” is used, the term “about” modifies both x and y unless the context otherwise specifies, and can be used interchangeably with the phrase “about x to about y”.

[0079] As used herein, the interchangeable terms “individual” or “subject” or “patient” mean any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates such as humans.

[0080] As used herein, the terms “comprise” (and any form of inclusion, such as “comprise”, “comprises”, and “comprised”), “have” (and any form of having, such as “have” and “has”), “include” (and any form of inclusion, such as “includes” and “include”), or “contains” (and any form of containing, such as “contains” and “contains”) are inclusive or open-ended and do not exclude additional, unlisted elements or method steps. Any step or composition using the transitional phrase “comprise” or “comprising” may also be described as being identical to that described by the transitional phrase “consisting of” or “consists”.

[0081] As used herein, the term “contact” means bringing two elements together in an in vitro or in vivo system. For example, “contacting” a virus or vector described herein with an individual or patient or cells includes administering a virus to an individual or patient, such as a human, and, for example, introducing a compound into a sample containing cells or a purified preparation containing cells.

[0082] As used herein, the terms “fusion” or “linkage” when referring to proteins with different domains or heterologous sequences mean that the protein domains are part of the same peptide chain and are linked to each other by peptide bonds or other covalent bonds. Domains or segments 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 are directly linked or fused to each other, or adapter sequences such as the glycine / serine sequences described herein link two domains together.

[0083] As used herein, "flexible stem" or "flexible stem protein" refers to a polypeptide containing a flexible region, which in some embodiments may be linked to an extracellular domain, a transmembrane domain, and / or an intracellular domain. The extracellular domain may be a target-binding domain, which may be referred to as "T".

[0084] In some implementations, a “mutant” or “variant” Fc protein is 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 otherwise requires.

[0085] "Disease" is a state of health in which the animal is unable to maintain homeostasis, and in which the animal's health continues to deteriorate if the disease is not treated. In contrast, an animal's "symptom" is a state of health in which the animal is able to maintain homeostasis, but in which the animal's health is worse than it would be without the symptom. A symptom, if left untreated, does not necessarily lead to a further decline in the animal's health.

[0086] The terms "effective amount" or "therapeutic effective amount" are used interchangeably herein and refer to the amount of a compound, formulation, material, or composition as described herein that effectively achieves a particular biological outcome or provides a therapeutic or preventative benefit. Such an outcome may include, but is not limited to, the amount of immune cell activation that, when administered to a mammal, results in a detectable level of immune cell activation compared to that detected in the absence of the composition. Immune responses can be readily assessed using a wide range of methods recognized in the art. Those skilled in the art will understand that the amount of the composition administered herein varies and can be readily determined based on many factors, such as the disease or symptom being treated, the age and health and physical condition of the mammal being treated, the severity of the disease, the specific compound administered, etc.

[0087] "Encoding" refers to the inherent characteristics of a specific nucleotide sequence in a polynucleotide (such as a gene, cDNA, or mRNA) for use as a template in biological processes to synthesize other polymers and macromolecules having defined nucleotide sequences (i.e., rRNA, tRNA, and mRNA) or defined amino acid sequences, and the resulting biological characteristics. Therefore, if the transcription and translation of the mRNA corresponding to a gene produces a protein in a cell or other biological system, then the gene encodes that protein. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in the sequence listing, and the non-coding strand, which serves as a template for transcription of the gene or cDNA, can be referred to as the protein or other product encoding that gene or cDNA.

[0088] "Expression vector" refers to a vector containing a recombinant polynucleotide that includes an expression control sequence operatively linked to a nucleotide sequence to be expressed. The expression vector contains sufficient cis-acting elements for expression; other elements for expression may be provided by a host cell or in an in vitro expression system. Expression vectors include all expression vectors known in the art, such as clomids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., Sendai virus, lentivirus, retrovirus, adenovirus, and adeno-associated virus) incorporating recombinant polynucleotides.

[0089] As used in this article, the phrase “ex vivo” in relation to transducing, transfecting or transforming cells means transducing, transfecting or transforming cells outside of a subject, i.e., removing cells from a subject prior to transducing, transfecting or transforming such cells.

[0090] As used herein, “identity” refers to the subunit sequence identity between two polymer molecules, such as two nucleic acid or amino acid molecules, or such as two polynucleotide or polypeptide molecules. Two amino acid sequences are identical when they have the same residue at the same position; for example, if each position in two polypeptide molecules is occupied by arginine, then they are identical at that position. The degree of identity, or similarity, between two amino acid or two nucleic acid sequences in an alignment is usually expressed as a percentage. Identity between two amino acid or two nucleic acid sequences is a direct function of the number of matching or identical positions; for example, if half of the positions in two sequences are identical, then the two sequences are 50% identical; if 90% of the positions (e.g., 9 out of 10) are matching or identical, then the two amino acid sequences are 90% identical.

[0091] "Substantially identical" means that the polypeptide or nucleic acid molecule exhibits at least 50% identity with 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 a sequence is at least 60%, 80%, or 85%, or 90%, 95%, or even 99% identical to the sequence used for comparison at the amino acid level or nucleic acid level. Other percentages of identity with respect to specific sequences are described herein.

[0092] Sequence identity can be measured / determined using sequence analysis software, such as the sequence analysis software package from Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, including the BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs. Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. Conserved substitutions typically include those within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In exemplary methods for determining the degree of identity, the BLAST program can be used, where a probability score between e3 and e100 indicates closely related sequences. In some embodiments, sequence identity is determined using BLAST with default settings.

[0093] For the purposes of the embodiments provided herein, compositions comprising various proteins may, in some cases, contain amino acid sequences that have sequence identity with the amino acid sequences disclosed herein. Therefore, in some embodiments, depending on the specific sequence, the degree of sequence identity with the SEQ ID NO disclosed herein is preferably greater than 50% (e.g., 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater). Other identity percentages are also provided herein in addition to these percentages. Identity between peptides can be determined using an affine gap search with parameters gap opening penalty (12) and gap extension penalty = 1, as implemented in the MPSRCH program (Oxford Molecular).

[0094] These proteins, compared to publicly available proteins, may include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conserved amino acid substitutions, where one amino acid is replaced by another amino acid with a related side chain. Genetically encoded amino acids are generally classified into four families: (1) acidic, i.e., aspartic acid and glutamic acid; (2) basic, i.e., lysine, arginine, and histidine; (3) nonpolar, i.e., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; and (4) uncharged polar, i.e., glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes collectively classified as aromatic amino acids. Generally, substitution of a single amino acid within these families does not significantly affect biological activity. Proteins may have one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) single amino acid deletions relative to publicly available protein sequences. In addition to the publicly available protein sequence, the protein may also contain one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) insertions (e.g., each of 1, 2, 3, 4, or 5 amino acids).

[0095] As used in this article, the phrase “in vivo” in relation to transducing, transfecting, or transforming cells means transducing, transfecting, or transforming such cells in a subject without removing the cells from the subject prior to transducing, transfecting, or transforming them.

[0096] "Separated" means altered or removed from its natural state. For example, nucleic acids or peptides that are naturally present in living organisms are not "separated," but the same nucleic acids or peptides that are partially or completely separated from their natural counterparts are "separated." Separated nucleic acids or proteins can exist in a substantially purified form or in non-natural environments, such as, for example, host cells.

[0097] As used herein, “lentivirus” refers to a genus of the family Retroviridae capable of infecting non-dividing cells. Non-limiting examples of lentiviruses are HIV, SIV, and FIV. Vectors or virus-like particles derived from lentiviruses can be used to transduce cells and deliver genes or other molecules, enabling their expression in cells either in vitro (ex vivo) or in vivo.

[0098] As used herein, the term "modified" refers to an altered state or structure of a molecule or cell as presented herein. Molecules can be modified in many ways, including chemically, structurally, and functionally, such as through mutation, substitution, insertion, or deletion (e.g., internal deletion truncation). Cells can be modified by introducing nucleic acids or expressing heterologous proteins.

[0099] As used herein, the term "modulation" means mediating an increase or decrease in a subject's response level compared to the response level of a subject in the absence of treatment or the compound, and / or compared to the response level of other subjects who are otherwise identical but untreated. This term includes interfering with and / or influencing natural signals or responses, thereby mediating a beneficial therapeutic response in a subject (such as a human).

[0100] Unless otherwise specified, "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate to each other and encode the same amino acid sequence. Phrases encoding protein or RNA may also contain introns, to the extent that nucleotide sequences encoding proteins may contain introns in some forms.

[0101] The term "oligonucleotide" usually refers to short polynucleotides. It should be understood that when a nucleotide sequence is represented by a DNA sequence (i.e., A, T, C, G), this also provides the corresponding RNA sequence (i.e., A, U, C, G) in which "U" replaces "T".

[0102] "Parenteral" administration of the composition includes techniques such as subcutaneous (sc), intravenous (iv), intramuscular (im), or intrasternal injection or infusion.

[0103] As used herein, the term "polynucleotide" is defined as a nucleotide chain. Furthermore, nucleic acids are polymers of nucleotides. Therefore, as used herein, the terms "nucleic acid" and "polynucleotide" are interchangeable. As used herein, polynucleotides include, but are not limited to, all nucleic acid sequences obtained by any method available in the art, including, but not limited to, recombinant methods, i.e., cloning nucleic acid sequences from recombinant libraries or cell genomes using cloning techniques and PCR, as well as synthetic methods.

[0104] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably and refer to compounds consisting of multiple amino acid residues covalently linked by peptide bonds. As used herein, the term refers to both short chains (which are also commonly referred to in the art as, for example, peptides, oligopeptides, and oligomers) and long chains (which are commonly referred to in the art as proteins, of which there are many types). “Polypeptide” includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, and so on. Polypeptides include natural peptides, recombinant peptides, synthetic peptides, or combinations thereof.

[0105] As used herein, the term "pseudotyped" or "pseudotyped viral particle" refers to a viral particle carrying a glycoprotein derived from another enveloped virus or a viral vector encoding an envelope glycoprotein from a virus different from its parent virus. Therefore, the host range of a vector particle can be expanded or altered depending on the type of cell surface receptor used by the glycoprotein. For example, viruses can be pseudotyped using VSV-G mutant proteins as described herein.

[0106] As used herein with respect to antibodies, the term "specifically binding" refers to an antibody that recognizes a specific antigen but substantially does not recognize or bind 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, this cross-species reactivity itself does not alter the antibody's specific classification. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, this cross-reactivity itself does not alter the antibody's specific classification. In some cases, the term "specifically binding" or "specifically binding" may be used in relation to the interaction of an antibody, protein, or peptide with a second chemical substance to mean that the interaction depends on the presence of a specific structure on the chemical substance (e.g., an antigenic determinant or epitope); for example, the antibody recognizes and binds to a specific protein structure, rather than a protein in general. If an antibody is specific for epitope "A," then in a reaction containing labeled "A" and an antibody, the presence of a molecule containing epitope A (or free, unlabeled A) will reduce the amount of labeled A that binds to the antibody. In some implementations, the targeting portions of viral particles containing mutant VSV-G proteins or other viral structural proteins used for pseudotypening of viruses, as described herein, can bind specifically to their targets.

[0107] The term "subject" includes living organisms, including those that can elicit an immune response (e.g., mammals). As used herein, a "subject" or "patient" can be a 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 rodents. In some embodiments, the subject is a human.

[0108] As used in this article, the term "therapeutic" refers to treatment and / or prevention. Therapeutic effects are achieved by suppressing, alleviating, or eradicating a disease state.

[0109] As used herein, the terms “transfected,” “transformed,” or “transduced” refer to the process of transferring or introducing exogenous nucleic acids into cells. “Transfected,” “transformed,” or “transduced” cells are cells that have been transfected, transformed, or transduced with exogenous nucleic acids. Cells include primary subject cells and their progeny. In some embodiments, transfection, transformation, or transduction occurs in vivo.

[0110] As used in this article, “treatment” means reducing the frequency or severity of at least one sign or symptom of a disease or condition experienced by the subject.

[0111] A "vector" is a composition of material containing isolated nucleic acids encoding proteins or peptides. Many 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, retroviral vectors, lentiviral vectors, etc.

[0112] "Carriers" or "delivery mediators" include viral particles, viruses, polylysine compounds, and liposomes that facilitate the transfer of nucleic acids into cells. Carriers or delivery mediators can also be used to deliver proteins or peptides into cells.

[0113] Scope: Throughout this disclosure, various aspects of the embodiments may be presented in a scope format. It should be understood that the scope format is for convenience and brevity only and should not be construed as an inflexible limitation. Therefore, the scope description should be considered as having specifically disclosed all possible sub-scopes and the individual values ​​within those scopes. For example, a scope such as 1 to 6 should be considered as having specifically disclosed sub-scopes such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and the individual numbers within those scopes, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the width of the scope. Unless otherwise expressly stated to the contrary, the disclosed scope also includes the endpoints of the scope.

[0114] Viral particle

[0115] In some implementations, viral particles comprising a heterologous viral glycoprotein, a targeting portion, and at least one nucleic acid molecule encoding a gene editing system are provided.

[0116] In some embodiments, the targeting portion comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stem portion. In some embodiments, S1 comprises a variant Fc protein, wherein the variant Fc protein comprises a transmembrane domain, such as, but not limited to, a CD8 or CD28 transmembrane domain. Thus, in some embodiments, the stem portion S1 comprises an N-terminal to C-terminal orientation (transmembrane domain) of the variant Fc. In some embodiments, the variant Fc protein comprises an effector mutation, wherein the effector mutation inhibits the interaction between the Fc protein and Fc-interacting proteins, such as FcγR, C1q, FcRβ, or FcRn.

[0117] In some embodiments, the S1 stem 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 comprises a variant of the sequence SEQ ID NO: 26 (IgG1 Fc), SEQ ID NO: 27 (IgG2 Fc), or SEQ ID NO: 28 (IgG4 Fc).

[0118] In some embodiments, the mutant Fc protein is the variant IgG1 Fc protein (SEQ ID NO: 26). In some embodiments, the variant IgG1 Fc protein comprises one or more mutations corresponding to those mutations selected from the group consisting of: L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26 according to the Kabat EU number index, as described in Edelman, GM et al., “The covalent structure of anentire 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. The mutations L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A and L235A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to N297A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to P329G of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, and P329G of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to I253A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H310A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H435A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to I253A, H310A, and H435A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26.

[0119] In some embodiments, the variant Fc protein comprising the variant IgG1 Fc protein comprises a truncated IgG1 Fc sequence. Truncating 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 comprising the variant IgG1 Fc protein comprises a truncated SEQ ID NO: 26. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of SEQ ID NO: 26. In some embodiments, truncating includes the deletion of amino acids from the N-terminus of SEQ ID NO: 26. In some embodiments, truncating includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 26. In some embodiments, the truncated SEQ ID NO: 26 comprises the amino acid sequence of SEQ ID NO: 81:

[0120] EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 81)

[0121] In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further contains one or more mutations corresponding to those 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 refer 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. The mutations L19A, L20A, N82A, P114G, I38A, H95A, and H220A of SEQ ID NO: 81 may or may not be present, and the 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 contains 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 contains 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 contains 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 contains 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 contains a mutation corresponding to I38A of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further contains a mutation corresponding to H95A of SEQ ID NO: 81. In some embodiments, the variant IgG1 Fc protein containing the amino acid sequence of SEQ ID NO: 81 further contains a mutation corresponding to 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 contains 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 contains 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 containing 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.

[0122] EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMASRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLAQDWLNGKEYKCKVSNKALGAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNAYTQKSLSLSPGK (SEQ ID NO: 82)

[0123] In some embodiments, the mutant Fc protein is the variant IgG2 Fc protein (SEQ ID NO: 27). In some embodiments, the variant IgG2 Fc protein comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A according to the Kabat EU number index, because those positions correspond to SEQ ID NO: 27. Any of the mutations N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 27 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the variant IgG2 Fc protein comprises a mutation corresponding to N297A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation corresponding to P329G of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises mutations corresponding to both N297A and P329G of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to I253A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to H310A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to H435A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains mutations corresponding to I253A, H310A, and H435A of SEQ ID NO: 27.

[0124] In some embodiments, the variant Fc protein comprising the variant IgG2 Fc protein comprises a truncated IgG2 Fc sequence. Truncating 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 comprising the variant IgG2 Fc protein comprises a truncated SEQ ID NO: 27. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of SEQ ID NO: 27. In some embodiments, truncating includes the deletion of amino acids from the N-terminus of SEQ ID NO: 27. In some embodiments, truncating includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 27.

[0125] In some embodiments, the mutant Fc protein is the variant IgG4 Fc protein (SEQ ID NO: 28). In some embodiments, the variant IgG4 Fc protein comprises one or more mutations selected from the group consisting of: S228P, L235E, N297A, P329G, I253A, H310A, and H435A according to the Kabat EU number index, because those positions correspond 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 the mutations may be combined in any combination. In some embodiments, the variant IgG4 Fc protein comprises the mutation corresponding to S228P of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises the mutation corresponding to L235E of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to N297A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to P329G of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains mutations corresponding to S228P, L235E, N297A, and P329G of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to I253A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to H310A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to H435A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains mutations corresponding to I253A, H310A, and H435A of SEQ ID NO: 28.

[0126] In some embodiments, the variant Fc protein comprising the variant IgG4 Fc protein comprises a truncated IgG4 Fc sequence. Truncating 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 comprising the variant IgG4 Fc protein comprises a truncated SEQ ID NO: 28. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of SEQ ID NO: 28. In some embodiments, truncating includes the deletion of amino acids from the N-terminus of SEQ ID NO: 28. In some embodiments, truncating includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 28.

[0127] In some embodiments, the stem portion S1 comprises a variant Fc protein given by formula L1-Fc-L2-X1, wherein L1 is a linker or is absent; Fc is a variant Fc protein; L2 is a linker or is absent; and X1 is a polypeptide comprising a transmembrane domain. Therefore, the targeting portion comprising formula T-S1 can also be given by formula T-L1-Fc-L2-X1, wherein T is a target-binding domain, L1 is a linker or is absent; Fc is a variant Fc protein; L2 is a linker or is absent; and X1 is a polypeptide comprising a transmembrane domain. Therefore, it should be understood that in some embodiments, the stem portion S1 can be given by formula L1-Fc-L2-X1. In some embodiments, the target-binding domain T is as provided herein. In some embodiments, the mutant Fc protein is as provided herein.

[0128] In some embodiments, L1 and L2 are each independently a peptide linker. In some embodiments, the peptide linker includes (GGGGA). n (SEQ ID NO: 54), (GGGGS) n (SEQ ID NO: 55), (EAAAK) n (SEQ ID NO: 73), A(EAAAK) n A (SEQ ID NO: 74), (XP) n(SEQ ID NO: 75) (where 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, wherein each n is independently 1-5. 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 not present. In some embodiments, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1-5. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 2. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 3. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 4. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 5. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently greater than 5. In some embodiments, L2 is absent. In some embodiments, L2 is (GGGGA).n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1-5. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 2. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 3. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 4. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO:55), where each n is independently 5. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n Where each n is independently greater than 5. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) and n is 1. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) and n is 3. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) and n is 4. In some implementations, L1 is (GGGGS). n (SEQ ID NO: 55) and n is 1. In some implementations, L1 is (GGGGS). n (SEQ ID NO: 55) and n is 2. In some implementations, L1 is (GGGGS). n(SEQ ID NO: 55) and n is 3. In some implementations, L1 is (GGGGS). n (SEQ ID NO: 55) and n is 4. In some embodiments, L1 is GSAGSAAGSGEF (SEQ ID NO: 56). In some embodiments, L1 is KESGSVSSEQLAQFRSLD (SEQ ID NO: 57). In some embodiments, L1 is EGKSSGSGSESKST (SEQ ID NO: 58).

[0129] In some implementations, X1 includes a formula ECD-T M -ICD polypeptides, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the targeting portion into the viral particle envelope, or it may not exist. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which may also be given by formula T-L1-Fc-L2-X1, may also be given by formula T-L1-Fc-L2-ECD-T. M -ICD is given, where T is the target-binding domain; L1 is the linker or absent; Fc is the variant Fc protein; L2 is the linker or absent; ECD is the extracellular domain or fragment of a cell surface protein, or absent; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the target portion into the viral particle envelope, or it may not exist. Therefore, it should be understood that in some embodiments, the stem portion S1 may be composed of the formula L1-Fc-L2-ECD-T M -ICD provides this information.

[0130] In some embodiments, the stem portion S1 does not contain a variant Fc region. In some embodiments, the stem portion S1 is given by formula L3-X1, where L3 is a flexible peptide linker and X1 is a polypeptide containing a transmembrane domain as provided herein. Therefore, in some embodiments, the targeting portion containing formula T-S1 may also be given by formula T-L3-X1, where T is a target-binding domain, L3 is a flexible peptide linker, and X1 is a polypeptide containing a transmembrane domain as provided herein. Therefore, it should be understood that in some embodiments, the stem portion S1 may be given by formula L3-X1. In some embodiments, the S1 stem portion is attached to the surface of the viral particle via a transmembrane domain.

[0131] In some embodiments, the flexible peptide linker L3 can be any flexible peptide linker. In some embodiments, L3 is selected from the group consisting of flexible linkers, including but not limited to (GGGGA).n (SEQ ID NO: 54), (GGGGS) n (SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESSKST (SEQ ID NO: 58), or any combination thereof, wherein each n is independently an integer selected from 1 to 4. In some embodiments, each n is independently an integer 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 (SEQ ID NO: 54) and n is 1. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L3 is (GGGGA). n (SEQ ID NO:54) and n is 3. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 4. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 5. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 6. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 7. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 8. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 9. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 10. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is greater than 10. In some implementations, L3 is (GGGGS). n(SEQ ID NO: 55) and n is 1. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 2. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 3. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 4. In some implementations, L3 is (GGGGS). n (SEQ ID NO:55) and n is 5. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 6. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 7. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 8. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 9. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 10. In some implementations, 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).

[0132] In some embodiments, the flexible peptide linker L3 can be any flexible peptide linker. In some embodiments, L3 is selected from the group consisting of flexible linkers, including but not limited to (GGGGA). n (SEQ ID NO: 54), (GGGGS) n(SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESSKST (SEQ ID NO: 58), or any combination thereof, 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 implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 3. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 4. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 1. In some implementations, L3 is (GGGGS). n (SEQ ID NO:55) and n is 2. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 3. In some implementations, 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).

[0133] In some implementations, L3 is selected from the group consisting of flexible joints, including but not limited to (GGGGA). n (SEQ ID NO:54), (GGGGS) n(SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESSKST (SEQ ID NO: 58), or any combination thereof, wherein each n is independently an integer 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 (SEQ ID NO: 54) and n is 1. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L3 is (GGGGA). n (SEQ ID NO:54) and n is 4. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 1. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 2. In some implementations, 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).

[0134] In some implementations, X1 includes a formula ECD-T M -ICD polypeptides, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the targeting portion into the viral particle envelope, or it is absent. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which may also be given by formula T-L3-X1, may also be given by formula T-L3-ECD-T. M -ICD is given, where T is the target-binding domain, L3 is the flexible peptide linker, and ECD is the extracellular domain of a cell surface protein or a fragment thereof, or it may not exist; T MIt is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the target portion into the viral particle envelope, or it may not exist. Therefore, it should be understood that in some embodiments, the stem portion S1 may be derived from formula L3-ECD-T. M -ICD provides this information.

[0135] In some embodiments, the ECD is absent. In some embodiments, the ECD can be any suitable extracellular domain or a fragment thereof. In some embodiments, the ECD originates from a different protein than the transmembrane domain. The ECD domain can be the entire ECD domain or a fragment thereof. In some embodiments, the ECD domain is a CD8 or CD28 ECD domain or a fragment thereof. In some embodiments, the ECD domain is a CD8 ECD domain or a fragment thereof. In some embodiments, the CD8 ECD domain comprises a polypeptide of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the CD8 ECD domain consists of or is substantially composed of a polypeptide of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain comprises a polypeptide of 25-45 amino acids in length. In some embodiments, the ECD comprises at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the peptide of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain is a CD28 ECD domain or a fragment thereof. In some embodiments, the CD28 ECD domain comprises a polypeptide of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the CD28 ECD domain is composed of or substantially composed of the polypeptide of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the ECD domain comprises a polypeptide of 25-45 amino acids in length. In some embodiments, the ECD contains at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the peptide of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO:60).

[0136] In some implementations, T M It can be any suitable transmembrane domain or fragment thereof. In some embodiments, T MThe structural domain is CD8 or CD28 T M A domain or a fragment thereof. In some implementations, T M The structural domain is CD8 T M A domain or a fragment thereof. In some implementations, CD8 T M The domain contains a polypeptide of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, CD8 T M The domain is composed of or substantially composed of a polypeptide of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M The polypeptide contains at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the peptide identical to that of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M The structural domain is CD28T M A domain or a fragment thereof. In some implementations, CD28 T M The domain comprises a polypeptide of the form FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, CD28 T M The domain is composed of or substantially composed of a polypeptide of the form FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, T M The polypeptide contains at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the same peptide as FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62).

[0137] In some implementations, T M The domain originates from the same protein as ECD. In some implementations, T M The domain originates from a protein different from ECD. In some implementations, T M The structural domain is CD8 or CD28 T M A structural domain or a fragment thereof, and the ECD structural domain is a CD8 or CD28 ECD structural domain or a fragment thereof. In some embodiments, T MThe peptide with the structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The peptide with the structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, T M The peptide with the structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62), and the ECD structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The peptides of FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62) have at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in terms of their structural domains, and the peptides of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60) have at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in terms of their ECD domains.

[0138] In some embodiments, the transmembrane domain is linked to the intracellular domain (ICD) of a cellular transmembrane protein or a fragment thereof. In some embodiments, the ICD is absent. In some embodiments, the ICD originates from T... M Proteins with the same or different domains. In some embodiments, the ICD contains an Env incorporation motif. An Env incorporation motif is a molecule, such as a polypeptide, that can facilitate protein incorporation into the viral envelope. A non-limiting example of an Env incorporation motif is a polypeptide containing the amino acid sequence NRVRQGYS (SEQ ID NO: 63). This is a non-limiting example, and other peptide sequences, such as, but not limited to, GGTETSQVAPA (SEQ ID NO: 64), can be used. In some embodiments, the Env incorporation motif contains the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 64, or a combination thereof. In some embodiments, the Env incorporation motif contains the amino acid sequence of SEQ ID NO: 63. In some embodiments, the Env incorporation motif contains the amino acid sequence of SEQ ID NO: 64.

[0139] In any of the embodiments described herein, the target-binding domain “T” is any polypeptide or polynucleotide that can be used to bind to the desired target. In some implementations, T is any polypeptide, polynucleotide, or fragment thereof that binds to: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR α, TCR β, TCR γ, TCRδ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; glycosylated CD43 epitopes expressed on acute leukemia or lymphoma but not on hematopoietic progenitor cells; glycosylated CD43 epitopes expressed on non-hematopoietic cancers; A kinase anchoring protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; anaplastic lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (TIP). 2) Anti-desmosome 1 (Dsg1) autoantibody, anti-desmosome 3 (Dsg3) autoantibody, B7H3 (CD276), biotin, bone marrow stromal cell antigen 2 (BST2), BST1 / CD157, cancer / testis antigen 1 (NY-ESO-1), cancer / testis antigen 2 (LAGE-la), carbonic anhydrase IX (CAIX), carcinoembryonic antigen (CEA), CCCTC binding factor (zinc finger protein)-like (BORIS or imprinted site regulator sibling factor), CCR4, CD5, CD19, CD20, CD22, CD24, CD30, CD32 (FCG) R2A), CD33, CD34, CD38, CD44v6, CD72, CD79a, CD79b, CD97, CD99, CD123, CD171, CD179a, CD179b-IGLll, CD200R, CD276 / B7H3, CD300 molecular-like family member f (CD300LF), CDH1-CD324, CDH6, CDH17, CDH19, chromosome X open reading frame 61 (CXORF61), sealing protein 6 (CLDN6), sealing protein 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-α), 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 2 containing mucin-like hormone receptor-like receptor (EMR2), mutated elongation factor 2 (ELF2M), liver glycoside B2, liver glycoside type A receptor 2 (EphA2), epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRviii), epithelial cell adhesion molecule (EPCAM), ERG, ETS translocation-variant gene 6 located on chromosome 12p (ETV6-AML), Fc fragment of IgA receptor (FCAR or CD89), Fc receptor-like 5 (FCRL5), fibroblast activation protein α (FAP) ), FITC, Fms-like tyrosine kinase 3 (FLT3), folate receptor α (FRa or FR1), folate receptor β (FRb), follicle-stimulating hormone receptor (FSHR), Fos-associated antigen 1, fucosylation-GM1, G protein-coupled receptor class C5 member D (GPRC5D), G protein-coupled receptor 20 (GPR20), GAD, ganglioside G2 (GD2), ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(ll)Cer), ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l- 4) bDGlcp(ll)Cer), GD3, GFRα4, glycoprotein 100 (gp100), phosphatidylinositol proteoglycan-3 (GPC3), gonadotropin receptor (CGHR or GR), GpA33, GpNMB, GPRC5D, guanylate cyclase C (GCC), mutant heat shock protein 70-2 (muthsp70-2), hepatitis A virus cell receptor 1 (HAVCR1), globoH glycosylneurase The hexasaccharide moiety of amide (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 (HPVE6), human papillomavirus E7 (HPV E7), human telomerase reverse transcriptase (hTERT), IgE, IL13Ra2, IL11Ra, immunoglobulin λ-like polypeptide 1 (IGLL1), influenza A hemagglutinin (HA), insulin-like growth factor 1 receptor (IGF-I receptor), interleukin-11 receptor α (IL-11Ra), interleukin-13 receptor subunit α-2 (IL-13Ra2 or CD213A2), intestinal carboxylesterase, KIT (CD117), KSHVK8.1, KSHV-gH, LAMP1, pod protein, 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, Livl, locus K 9 (LY6K), low-conductivity chloride channel, lymphocyte antigen 6 complex, lymphocyte antigen 75 (LY75), lymphocyte-specific protein tyrosine kinase (LCK), breast differentiation antigen (NY-BR-1), T-cell recognized melanoma antigen 1 (MelanA or MARTI), melanoma-associated antigen 1 (MAGE-A1), melanoma cancer testis antigen-1 (MAD-CT-1), melanoma cancer testis antigen-2 (MAD-CT-2), melanoma cell apoptosis inhibitor (ML-IAP), mesothelin, MPL, cell surface-associated mucin 1 (MUC1), N-acetylglucosamine transferase V (NA17), connexin-4, neural cell adhesion molecule (NCAM), NKG2D, NYBR1, O-acetyl-GD2 ganglioside (OAcGD2), olfactory receptor 51E2 (OR51E2), from breakpoint The following proteins are listed: a fusion protein composed of the cluster region (BCR) and Abelson murine leukemia virus oncogene homolog 1 (Abl) (bcr-abl); a P53 mutant; pairing box proteins Pax-3 (PAX3), Pax-5 (PAX5), pan-connecting protein 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor β (PDGFR-β); polysialic acid; proaporthetin-binding protein sp32 (OY-TES1); prostate enzymes; prostate cancer tumor antigen-1 (PCTA-1 or galactohemagglutinin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA); prostate acid phosphatase (PAP); Prostein; serine protease 21 (Testisin or PRSS21); proteasome macropain subunit β9 (LMP2); PTK7; and Ras.G12V, Ras homolog family member C (RhoC), Rat sarcoma (Ras) mutant, receptor for advanced glycation end products (RAGE-1), receptor tyrosine kinase-like orphan receptor 1 (ROR1), receptor tyrosine protein kinase ERBB2 or Her-22 / neu, renal ubiquitin 1 (RU1), renal ubiquitin 2 (RU2), sarcoma translocation breakpoint, serine 2 (TMPRSS2) ETS fusion gene, sialic acid Lewis adhesion molecule (sLe), SLAMF4, SLAMF6, Slea (CA19.9 or sialic acid Lewis antigen), spermin 17 (SPA17), T cell recognized squamous cell carcinoma antigen 3 (SART3), stage-specific embryonic antigen-4 (SSEA-4), STEAP1, survival protein, synovial sarcoma X breakpoint 2 (SSX2), TCR γ-substituted reading frame protein (TARP), TCR-β1 chain, TCR-β2 chain, TCR-δ chain, TCR-γ chain, TCRγ-δ, telomerase, TGFβR2, antigen recognized by TNT antibody, thyroid-stimulating hormone receptor (TSHR), Timol / HVCR1, tissue factor 1 (TF1), Tn 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 associated (TEM7R), tumor protein p53 (p53), tumor-associated glycoprotein 72 (TAG72), tyrosinase, tyrosinase-associated protein 2 (TRP-2), urolytic protein 2 (UPK2), vascular endothelial growth factor receptor 2 (VEGFR2), V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN), Wilms tumor 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. It should be understood that, in the context of this disclosure, "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 any isotype of antibody, antibody fragment (including but not limited to Fab, Fv, scFv, and Fd fragments), chimeric antibody, humanized antibody, single-chain antibody (scAb), single-domain antibody (dAb), single-domain heavy chain antibody, single-domain light chain antibody, bispecific antibody, multispecific antibody, and fusion protein 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 antibody, 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.

[0140] In some embodiments, the viral particles provided herein are pseudotyped viral particles. In some embodiments, viral particles are pseudotyped using viral glycoproteins of viruses from the Paramyxoviridae family. In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of the morbillivirus genus (such as Measlesvirus). In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Measlesvirus. In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Henipavirus (such as Nipah virus, Cedar virus, or Hendra virus). In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Nipah virus. In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Nipah virus. In some embodiments, peptides or antibodies as provided herein are linked to envelope glycoproteins G or H of viruses from the Paramyxoviridae family via a linker. In some implementations, the viruses of the Paramyxoviridae family are those of the Measlesvirus genus, such as measles virus. In other implementations, the viruses of the Paramyxoviridae family are Hennipa viruses, such as Nipah virus, Cedar virus, or Hendra virus.

[0141] As provided herein, viruses can be pseudotyped using the VSV-G protein (wild type or a mutant thereof). Without being bound by any particular theory, a mutant VSV-G protein containing a mutation at position 182, which can be used to pseudotype viruses (e.g., lentiviruses) when the virus contains a target region, can be used to pseudotype the virus and transduce cells. This mutation inhibits or reduces the affinity of VSV-G for its natural 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 target cells.

[0142] In some embodiments, a VSV-G protein comprising a mutation at position 198 compared to SEQ ID NO: 1 or a mutation at position 182 compared to SEQ ID NO: 2 is provided. SEQ ID NO: 1 is the full-length protein, and SEQ ID NO: 2 is the extracellular domain of the VSV-G protein. The 16-mer signal peptide of MKCLLYLAFLFIGVNC (SEQ ID NO: 65) as shown at the N-terminus of SEQ ID NO: 1 is cleaved, leaving the protein of SEQ ID NO: 2. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 2, it should be understood that the mutation also occurs in the context of SEQ ID NO: 1 containing the leader sequence, and will therefore be a position number 16 positions higher than that described in 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 implementations, the mutation is the I182E mutation compared to SEQ ID NO: 2.

[0143] In some embodiments, a VSV-G protein comprising a mutation at position 198 compared to SEQ ID NO: 10 or a mutation at position 182 compared to SEQ ID NO: 11 is provided. SEQ ID NO: 10 is the full-length protein, and SEQ ID NO: 11 is the extracellular domain of the VSV-G protein. The 16-mer signal peptide of MLSYLIFALVVSPILG (SEQ ID NO: 66) as shown at the N-terminus of SEQ ID NO: 10 is cleaved, leaving the protein of SEQ ID NO: 11. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 11, it should be understood that the mutation also occurs in the context of SEQ ID NO: 10 containing the leader sequence, and will therefore be a position number 16 positions higher than that described in 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 implementations, the mutation is the T182E mutation compared to SEQ ID NO: 11.

[0144] In some embodiments, a VSV-G protein comprising a mutation at position 198 compared to SEQ ID NO: 12 or a mutation at position 182 compared to SEQ ID NO: 13 is provided. SEQ ID NO: 12 is the full-length protein, and SEQ ID NO: 13 is the extracellular domain 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. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 13, it should be understood that the mutation also occurs in the context of SEQ ID NO: 12 containing the leader sequence, and will therefore be a position number 16 positions higher than that described in 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 implementations, the mutation is the A182E mutation compared to SEQ ID NO: 13.

[0145] In some embodiments, a VSV-G protein comprising a mutation at position 203 compared to SEQ ID NO: 14 or a mutation at position 182 compared to SEQ ID NO: 15 is provided. SEQ ID NO: 14 is the full-length protein, and SEQ ID NO: 15 is the extracellular domain of the VSV-G protein. The 21-mer signal peptide of MKMKMVIAGLILCIGILPAIG (SEQ ID NO: 68) as shown at the N-terminus of SEQ ID NO: 14 is cleaved, leaving the protein of SEQ ID NO: 15. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 15, it should be understood that the mutation also occurs in the context of SEQ ID NO: 14 containing the leader sequence, and will therefore be a position number 21 more than the position described in 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 implementations, the mutation is the V182E mutation compared to SEQ ID NO: 15.

[0146] In some embodiments, a VSV-G protein comprising a mutation at position 199 compared to SEQ ID NO: 16 or a mutation at position 182 compared to SEQ ID NO: 17 is provided. SEQ ID NO: 16 is the full-length protein, and SEQ ID NO: 17 is the extracellular domain of the VSV-G protein. The 17-mer signal peptide of MTPAFILCMLLAGSSWA (SEQ ID NO: 69), as shown at the N-terminus of SEQ ID NO: 16, is cleaved, leaving the protein of SEQ ID NO: 17. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 17, it should be understood that the mutation also occurs in the context of SEQ ID NO: 16 containing the leader sequence, and will therefore be a position number 17 more than the position described in 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 implementations, the mutation is the V182E mutation compared to SEQ ID NO: 17.

[0147] In some embodiments, a VSV-G protein comprising a mutation at position 199 compared to SEQ ID NO: 18 or a mutation at position 182 compared to SEQ ID NO: 19 is provided. SEQ ID NO: 18 is the full-length protein, and SEQ ID NO: 19 is the extracellular domain 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. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 19, it should be understood that the mutation also occurs in the context of SEQ ID NO: 18 containing the leader sequence, and will therefore be a position number 17 positions higher than that described in 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 implementations, the mutation is the V182E mutation compared to SEQ ID NO: 19.

[0148] In some embodiments, a VSV-G protein comprising a mutation at position 199 compared to SEQ ID NO: 20 or a mutation at position 182 compared to SEQ ID NO: 21 is provided. SEQ ID NO: 20 is the full-length protein, and SEQ ID NO: 21 is the extracellular domain of the VSV-G protein. The 17-mer signal peptide of MLVLYLLLSLLALGAQC (SEQ ID NO: 71), as shown at the N-terminus of SEQ ID NO: 20, is cleaved, leaving the protein of SEQ ID NO: 21. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 21, it should be understood that the mutation also occurs in the context of SEQ ID NO: 20 containing the leader sequence, and will therefore be a position number 17 positions higher than that described in 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 implementations, the mutation is the I182E mutation compared to SEQ ID NO: 21.

[0149] As used herein, when a peptide is considered to have a mutation compared to a reference sequence, this comparison is based on alignment performed using software such as BlastP, ClustalW, or ClustalOmega with default parameters. For example, position 182 can be found in SEQ ID NO: 2, and is also related to... Figure 3 Compare with other strains shown. Figure 3 Clustal alignments of the wild-type sequences of the extracellular domain of the VSV-G protein from various strains are shown. Bold and underlined residues are those aligned to position 182 of SEQ ID NO: 2 for each strain. SEQ ID NO: 2 refers to the extracellular domain of the VSV-G protein from the Indiana strain. SEQ ID NO: 11 refers to the extracellular domain of the VSV-G protein from the New Jersey strain. SEQ ID NO: 13 refers to the extracellular domain of the VSV-G protein from the Malabar strain. SEQ ID NO: 15 refers to the extracellular domain of the VSV-G protein from the Carajá strain. SEQ ID NO: 17 refers to the extracellular domain of the VSV-G protein from the Alagoa strain. SEQ ID NO: 19 refers to the extracellular domain of the VSV-G protein from the Cocal strain. SEQ ID NO: 21 refers to the extracellular domain of the VSV-G protein from the Morton strain. Therefore, residues that are compared with residue 182 of SEQ ID NO: 2 can also be mutated as provided herein.

[0150] 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.

[0151] In some embodiments, the mutation at position 182 compared to SEQ ID NO: 2 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 11 is T182S, T182H, T182Q, or T182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 13 is A182S, A182H, A182T, A182Q, or A182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 15 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 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.

[0152] Although the mutation may be described with reference to SEQ ID NO: 1 or SEQ ID NO: 2 (which is the VSV-G protein from the Indiana strain), the mutation can also be used in other strains of the VSV-G protein. For example, the mutation can be made in the New Jersey strain of VSV-G, the Malaba strain of VSV-G, the Carajás strain of VSV-G, the Alagoa strain of VSV-G, the Cocal strain of VSV-G, or the Morton strain of VSV-G. In some embodiments, the sequence of each strain is as provided herein. Examples of these strains 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 extracellular domains of the New Jersey strain of VSV-G are SEQ ID NO:10 and SEQ ID NO:11, respectively; the wild-type full-length or extracellular domains of the Malaba strain of VSV-G are SEQ ID NO:12 and SEQ ID NO:13, respectively; the wild-type full-length or extracellular domains of the Carajás strain of VSV-G are SEQ ID NO:14 and SEQ ID NO:15, respectively; the wild-type full-length or extracellular domains of the Alagoa strain of VSV-G are SEQ ID NO:16 and SEQ ID NO:17, respectively; the wild-type full-length or extracellular domains of the Cocal strain of VSV-G are SEQ ID NO:18 and SEQ ID NO:19, respectively; or the wild-type full-length or extracellular domains of the Morton strain of VSV-G are SEQ ID NO:20 and SEQ ID NO:21, respectively.

[0153] The VSV-G protein containing the mutation at position 182 compared to SEQ ID NO: 2 may also contain other mutations, such as those described in U.S. Patent Application Publication No. 20200216502, the entire contents of which are incorporated herein by reference. For example, the VSV-G protein may contain mutations at positions corresponding to positions 8, 47, 209, and / or 354 of SEQ ID NO: 2.

[0154] In some embodiments, the substitution at position 8 is made by any amino acid other than Y that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 209 is made by any amino acid other than H that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 47 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 354 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2.

[0155] In some embodiments, the replacement is located at position 47 or at position 354, or at both positions 47 and 354, and is replaced by A, G, F, or Q. In some embodiments, the replacement is A or Q.

[0156] In some implementations, the substitution at position 8 is alanine, i.e., H8A.

[0157] In some implementations, the substitution at position 47 is Q or N, i.e., K47Q or K47N.

[0158] In some embodiments, the protein contains a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0159] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 2 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 to SEQ ID NO: 2 (or, if a full-length protein is used, to SEQ ID NO: 1). In some embodiments, the polypeptide contains an I182D or I182E mutation. In some embodiments, the VSV-G protein contains an I182S, I182H, I182T, I182Q, or I182N mutation.

[0160] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 11 contains the 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 to SEQ ID NO: 11 (or, if using a full-length protein, SEQ ID NO: 10). In some embodiments, the polypeptide contains a T182D or T182E mutation. In some embodiments, the VSV-G protein contains a T182S, T182H, T182Q, or T182N mutation.

[0161] In some embodiments, a protein containing 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 to SEQ ID NO: 13 (or, if using a full-length protein, to SEQ ID NO: 12). In some embodiments, the polypeptide contains an A182D or A182E mutation. In some embodiments, the VSV-G protein contains an A182S, A182H, A182T, A182Q, or A182N mutation.

[0162] In some embodiments, a protein containing 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 to SEQ ID NO: 15 (or, if using a full-length protein, SEQ ID NO: 14). In some embodiments, the polypeptide contains a V182D or V182E mutation. In some embodiments, the VSV-G protein contains a V182S, V182H, V182T, V182Q, or V182N mutation.

[0163] In some embodiments, a protein containing 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 to SEQ ID NO: 17 (or, if using a full-length protein, SEQ ID NO: 16). In some embodiments, the polypeptide contains a V182D or V182E mutation. In some embodiments, the VSV-G protein contains a V182S, V182H, V182T, V182Q, or V182N mutation.

[0164] In some embodiments, a protein containing 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 to SEQ ID NO: 19 (or, if using a full-length protein, to SEQ ID NO: 18). In some embodiments, the polypeptide contains a V182D or V182E mutation. In some embodiments, the VSV-G protein contains a V182S, V182H, V182T, V182Q, or V182N mutation.

[0165] In some embodiments, a protein containing 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 to SEQ ID NO: 21 (or, if using a full-length protein, SEQ ID NO: 20). In some embodiments, the polypeptide contains an I182D or I182E mutation. In some embodiments, the VSV-G protein contains an I182S, I182H, I182T, I182Q, or I182N mutation.

[0166] Viral glycoprotein

[0167] The mutant VSV-G protein can be used, for example, for pseudotypening viruses (such as, but not limited to, lentiviruses). Therefore, in some embodiments, viral particles comprising the mutant VSV-G protein as provided herein are provided. In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 198 compared to SEQ ID NO: 1. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 2 contains the 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 to SEQ ID NO: 2 (or, if using the full-length protein, to SEQ ID NO: 1). In some embodiments, the peptide contains an I182D or I182E mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein contains an I182S, I182H, I182T, I182Q, or I182N mutation.

[0168] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 198 compared to SEQ ID NO: 10. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 11 contains the 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 to SEQ ID NO: 11 (or, if using the full-length protein, to SEQ ID NO: 10). In some embodiments, the polypeptide contains a T182D or T182E mutation compared to SEQ ID NO: 11. In some implementations, the VSV-G protein contains T182S, T182H, T182Q, or T182N mutations.

[0169] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 198 compared to SEQ ID NO: 12. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 13 contains the 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 to SEQ ID NO: 13 (or, if using the full-length protein, to SEQ ID NO: 12). In some embodiments, the polypeptide contains an A182D or A182E mutation compared to SEQ ID NO: 13. In some implementations, the VSV-G protein contains mutations of A182S, A182H, A182T, A182Q, or A182N.

[0170] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 203 compared to SEQ ID NO: 14. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 15 contains the 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 to SEQ ID NO: 15. In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 15. In some implementations, the VSV-G protein contains mutations of V182S, V182H, V182T, V182Q, or V182N.

[0171] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 199 compared to SEQ ID NO: 16. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 17 contains the 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 to SEQ ID NO: 17. In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 17. In some implementations, the VSV-G protein contains mutations of V182S, V182H, V182T, V182Q, or V182N.

[0172] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 199 compared to SEQ ID NO: 18. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 19 contains the 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 to SEQ ID NO: 19 (or, if using the full-length protein, to SEQ ID NO: 18). In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 19. In some implementations, the VSV-G protein contains mutations of V182S, V182H, V182T, V182Q, or V182N.

[0173] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 199 compared to SEQ ID NO: 20. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 21 contains the 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 to SEQ ID NO: 21 (or, if using the full-length protein, to SEQ ID NO: 20). In some embodiments, the polypeptide contains an I182D or I182E mutation compared to SEQ ID NO: 21. In some implementations, the VSV-G protein contains I182S, I182H, I182T, I182Q, or I182N mutations.

[0174] In some embodiments, the VSV-G protein further comprises mutations at positions corresponding to positions 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 comprises a substitution at position I182 of SEQ ID NO: 2 and at least one of T214 and T352. In some embodiments, the VSV-G polypeptide comprises substitutions at positions I182, T214, and T352 of SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises a mutation corresponding to the T214N mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises a mutation corresponding to the T352A mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises both the T214N and T352A mutations compared to SEQ ID NO: 2. These mutations can be combined with any other mutations as provided herein. In some embodiments, the T214N and / or T352A mutations are combined with the I182E or I182D mutations. In some embodiments, the VSV-G protein comprises the amino acid sequences of SEQ ID NO: 22 and SEQ ID NO: 23, which combine I182D or I182E with the T214N and T352A mutations, respectively. These sequences are also illustrated below with the leader sequences that are removed during protein processing.

[0175] VSV-G protein with I196E, T230N, and T368A mutations (with leader sequence and adjusted numbering)

[0176] MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(SEQ ID NO:24)

[0177] VSV-G protein with I182E, T214N, and T352A mutations (without leader sequence)

[0178] KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(SEQ ID NO: 22)

[0179] VSV-G protein with I196E, T230N, and T368A mutations (with leader sequence and adjusted numbering)

[0180] MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(SEQ ID NO:25)

[0181] VSV-G protein with I182E, T214N, and T352A mutations (without leader sequence)

[0182] KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEA VIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAAR FPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVL RTSSGYKFPLYMIGGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 23)

[0183] In some embodiments, 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. In some embodiments, the VSV-G polypeptide comprises the sequence of SEQ ID NO: 22. In some embodiments, the VSV-G polypeptide comprises the sequence of SEQ ID NO: 23. In some embodiments, the VSV-G polypeptide comprises the sequence of SEQ ID NO: 24. In some embodiments, the VSV-G polypeptide comprises the sequence of SEQ ID NO: 25. It should be understood that the VSV-G sequences of SEQ ID NO: 22-25 are exemplary and are not intended to be limiting in any way. Other strains of the VSV-G protein described herein may also contain mutations corresponding to T214N and / or T352A in SEQ ID NO: 2 and as shown in SEQ ID NO: 22 and SEQ ID NO: 23. Such additional sequences are within the scope of this disclosure.

[0184] In some embodiments, the composition comprises mutations as described in Hwang et al., Gene Ther, Aug 2013; 20(8):807-15. (Epub, Jan 31, 2013), which is incorporated herein by reference in its entirety. For example, mutations may be at positions 230, 368, 66, and / or 162 corresponding to SEQ ID NO: 1. These positions would be 16 fewer than those in SEQ ID NO: 2 when the leader sequence is removed. In some embodiments, mutations at those positions are, for example, T230N, T368A, K66T, S162T, or any combination thereof. In some embodiments, the VSV-G protein comprises the T230N and T368A mutations. In some embodiments, the VSV-G polypeptide comprises K66T, S162T, T230N, and T368A. These positions are those corresponding to the positions in the full-length protein (SEQ ID NO: 1). In some embodiments, the VSV-G protein comprises the T230N mutation, the T368A mutation, the K66T mutation, the S162T mutation, or any combination thereof. In some embodiments, in addition to the mutation corresponding to position 182 of SEQ ID NO: 2, the VSV-G protein also comprises one or more mutations, such as those described in U.S. Patent Application Publication No. 20200216502, the entire contents of which are incorporated herein by reference. For example, the VSV-G protein may also comprise mutations at positions corresponding to positions 8, 47, 209, and / or 354 of SEQ ID NO: 2.

[0185] In some embodiments, the substitution at position 8 is made by any amino acid other than Y that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 209 is made by any amino acid other than H that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 47 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 354 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution is located at position 47 or position 354, or at both positions 47 and 354, and is substituted by 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.

[0186] In addition, in some implementations, other viral structural proteins can be used to pseudotype the virus instead of the VSV-G protein or its mutants.

[0187] For example, viral particles can be pseudotyped using the carp viremia virus G (SVCV-G) protein, and cells can be transduced when the virus contains a targeting portion. In some embodiments, the provided carp viremia virus G protein can be used to transduce target cells and deliver heterologous molecules to target cells. In some embodiments, a carp viremia virus G (SVCV-G) protein comprising SEQ ID NO: 52 is provided. SEQ ID NO: 52 is the full-length protein, and SEQ ID NO: 53 is the extracellular domain of the carp viremia virus G protein with the N-terminal signal peptide removed. Therefore, in some embodiments, the protein comprises the amino acid sequence of SEQ ID NO: 53. The carp viremia virus G protein can be used, for example, to pseudotype viruses (such as, but not limited to, lentiviruses). Therefore, in some embodiments, viral particles comprising the carp viremia virus G protein as provided herein are provided. In some embodiments, the viral particle contains a carp spring viremia virus G protein containing 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% of the same sequence as SEQ ID NO: 52 or SEQ ID NO: 53.

[0188] Sequence of Spring viremia of carp virus-G (SEQ ID NO: 52 - with leader sequence)

[0189] MSIISYIAFLLLIDSNLGIPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDLGFPPQSCGWASVTTVSNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWVEKTAGTLTTIHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELDGNIYQGFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0190] Sequence of Spring viremia of carp virus-G (SEQ ID NO: 53 - without leader sequence) :

[0191] IPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDLGFPPQSCGWASVTTV SNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWVEKTAGTLTT IHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELDGNIYQ GFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0192] Gene editing system

[0193] In some embodiments, the viral particles contain at least one nucleic acid molecule encoding a gene editing system. The term "gene editing system" refers to some or all of the components necessary for a functional gene editing complex. For example, a CRISPR / Cas9 gene editing system may contain guide RNA or a single guide RNA and the Cas9 protein. In another embodiment, a CRISPR / Cas9 gene editing system may contain one or more nucleic acid molecules encoding a single guide RNA and the Cas9 protein. In other embodiments, the gene editing system may include a zinc finger nuclease system, a transcription activator-like effector nuclease (TALEN) system, or a broad range of nuclease systems.

[0194] 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 TALEN. In some embodiments, the gene editing system is a broad-spectrum nuclease. In some embodiments, the gene editing system is a gene product regulating nucleic acid molecule. The gene product regulating the nucleic acid molecule is known in the art and any such product can be used. In some embodiments, the gene product regulating nucleic acid molecule is selected from, but is not limited to, siRNA, piRNA, miRNA, RNAi, mRNA, shRNA, and antisense RNA. In some embodiments, the gene product regulating nucleic acid molecule is siRNA. In some embodiments, the gene product regulating nucleic acid molecule is piRNA. In some embodiments, the gene product regulating nucleic acid molecule is miRNA. In some embodiments, the gene product regulating nucleic acid molecule is RNAi. In some embodiments, the gene product regulating nucleic acid molecule is mRNA. In some embodiments, the gene product regulating nucleic acid molecule is shRNA. In some embodiments, the gene product regulating nucleic acid molecule is antisense RNA.

[0195] In some embodiments, the gene editing system may be a CRISPR-Cas system. The CRISPR-Cas system may contain a CRISPR system guide or a polynucleotide encoding the guide. The guide directs the CRISPR complex to bind sequence-specifically to a target sequence at a genomic locus of the cell to be engineered. The target sequence may be modified by the CRISPR-Cas system. Alternatively or otherwise, the CRISPR-Cas system may contain a CRISPR-Cas protein or a polynucleotide encoding a CRISPR protein. In some cases, the CRISPR-Cas system may also contain a template or a polynucleotide encoding the template. The template may contain a nucleic acid sequence capable of modifying 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, but is 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 type V 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, but is 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 comprises dCas linked to cytidine deaminase or adenosine deaminase.In some embodiments, the CRISPR-Cas base editing system comprises dCas linked to cytidine deaminase. In some embodiments, the CRISPR-Cas base editing system comprises dCas linked to adenosine deaminase. In some embodiments, the CRISPR-Cas system is a lead editing system. In some embodiments, the CRISPR-Cas lead editing system comprises CRISPRa, CRISPRi, Cas3, or CasMINI proteins. In some embodiments, the CRISPR-Cas lead editing system comprises CRISPRa. In some embodiments, the CRISPR-Cas lead editing system comprises CRISPRi. In some embodiments, the CRISPR-Cas lead editing system comprises Cas3. In some embodiments, the CRISPR-Cas lead editing system comprises the CasMINI protein.

[0196] In some implementations, the gene editing system is a TALEN system. For example, a TALEN system may comprise a transcription activator-like effector nuclease (TALEN), an engineered variant thereof, or a nucleic acid encoding it.

[0197] In some implementations, the gene editing system is a ZFN system. For example, a ZFN system may comprise 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 with a DNA-cutting domain, which can be engineered to target a desired DNA sequence.

[0198] In some implementations, the gene editing system is a large-scale nuclease system. For example, a large-scale nuclease system may comprise a large-scale nuclease, an engineered variant thereof, or a nucleic acid encoding it. A large-scale nuclease may be a deoxyribonuclease characterized by a large recognition site (a double-stranded DNA sequence of 12 to 40 base pairs).

[0199] The term "endogenous" is used herein to indicate that a molecule is natural for a particular cell, cell population, cell type, tissue, or organism. For example, an endogenous gene is a gene that is natural for a particular cell, cell population, cell type, tissue, or organism. In some embodiments, a target nucleic acid molecule is edited, wherein the target nucleic acid molecule is an endogenous gene.

[0200] Targeting moiety

[0201] In some embodiments, the viral particle includes a targeting portion having the formula T-S1, where T is a target-binding domain and S1 is a stem portion. The targeting portion can be used to target viral particles containing mutant VSV-G or SVCV-G proteins to cells expressing a target to which the targeting portion binds. In some embodiments, the target-binding domain is an antibody, scFv antibody, antigen-binding domain, an ankyrin repeat sequence (e.g., DARPIN), a VHH domain antibody, a nanobody, a single-domain antibody, an 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 flexible 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 targeting portion attaches (fused to or linked to) the envelope glycoprotein G or H of a virus belonging to the Paramyxoviridae family, such as measlesvirus, or Hennipa virus, such as Nipah virus, cedar virus, or Hendra virus. In some embodiments, the targeting portion may attach (fused to or linked to) the glycoprotein of a virus belonging to the Rhabdoviridae family, such as vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), parainfluenza virus, fall armyworm rhabdovirus isolate Sf G, Drosophila sigmavirus 10A, Wuhan insect virus 7, perch virus, or carp spring viremia virus. In some embodiments, the VSV protein is a mutant protein, such as those described herein. In some implementations, the target portion is attached to the glycoprotein of viruses of the Filoviridae family (such as Ebola virus) or viruses of the Arenaviridae family (such as Machupo virus).

[0202] In some implementations, the target binding domain is scFv. In some implementations, the target binding domain is a single-domain antibody. In some implementations, the target binding domain is VHH.

[0203] In some implementations, the targeting portion binds to the following: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR α, TCR β, TCR γ, TCR δ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; glycosylated CD43 epitopes expressed on acute leukemia or lymphoma but not on hematopoietic progenitor cells; glycosylated CD43 epitopes expressed on non-hematopoietic cancers; A kinase anchoring protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; anaplastic lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie 2) Anti-desmosome mucin 1 (Dsg1) autoantibody; anti-desmosome mucin 3 (Dsg3) autoantibody; B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testis antigen 1 (NY-ESO-1); cancer / testis antigen 2 (LAGE-la); carbonic anhydrase IX (CAIX); carcinoembryonic antigen (CEA); CCCTC binding factor (zinc finger protein)-like (BORIS or imprinted site regulator sibling factor); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCG) R2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLll; CD200R; CD276 / B7H3; CD300 molecular-like family member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19; Chromosome X open reading frame 61 (CXORF61); Sealing protein 6 (CLDN6); Sealing protein 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-α); 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 2 containing mucin-like hormone receptor-like receptor (EMR2); Elongation factor 2 mutant (ELF2M); Hepatin B2; Hepatin A receptor 2 (EphA2)Epidermal growth factor receptor (EGFR); Epidermal growth factor receptor variant III (EGFRviii); Epithelial cell adhesion molecule (EPCAM); ERG; ETS translocation variant gene 6 located on chromosome 12p (ETV6-AML); Fc fragment of IgA receptor (FCAR or CD89); Fc receptor-like 5 (FCRL5); Fibroblast activation protein α (FAP); FITC; Fms-like tyrosine kinase 3 (FLT3); Folate receptor α (FRa or FR1); Folate receptor β (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-associated antigen 1; Fucosyl-GM1; G protein-coupled receptor class C5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(ll)Cer); Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l- 4) bDGlcp(ll)Cer); GD3; GFRα4; Glycoprotein 100 (gplOO); Phosphatidylinositol proteoglycan-3 (GPC3); Gonadotropin receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylate cyclase C (GCC); Mutant heat shock protein 70-2 (mut hsp70-2); Hepatitis A virus cell receptor 1 (HAVCR1); GloboH glycosylceramide hexasaccharide moiety (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 λ-like polypeptide 1 (IGLL1); Influenza A hemagglutinin (HA); Insulin-like growth factor 1 receptor (IGF-I receptor); Interleukin 11 receptor α (IL-11Ra); Interleukin 13 receptor subunit α-2 (IL-13Ra2 or CD213A2); Intestinal carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; pod protein; 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; Livl; Locus K9 (LY6K); Low-conductivity chloride channel;Lymphocyte antigen 6 complex; Lymphocyte antigen 75 (LY75); Lymphocyte-specific protein tyrosine kinase (LCK); Breast differentiation antigen (NY-BR-1); T-cell recognized melanoma antigen 1 (MelanA or MARTI); Melanoma-associated antigen 1 (MAGE-A1); Melanoma cancer testis antigen-1 (MAD-CT-1); Melanoma cancer testis antigen-2 (MAD-CT-2); Melanoma cell apoptosis inhibitor (ML-IAP); Mesothelin; MPL; Cell surface-associated mucin 1 (MUC1); N-acetylglucosamine transferase V (NA17); Ligin-4; Neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acetyl-GD2 ganglioside (OAcGD2); Olfactory receptor 51E2 (OR51E2); Composed of breakpoint cluster region (BCR) and A Oncogene fusion protein composed of Belson murine leukemia virus oncogene homolog 1 (Abl) (bcr-abl); P53 mutant; Pax-3 (PAX3); Pax-5 (PAX5); pan-connecting protein 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor β (PDGFR-β); polysialic acid; proapocrine-binding protein sp32 (OY-TES1); prostate enzymes; prostate cancer tumor antigen-1 (PCTA-1 or galactohemagglutinin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA); prostate acid phosphatase (PAP); Prostein; serine protease 21 (Testisin or PRSS21); proteasome (Prosome Macropain subunit β9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); Rat sarcoma (Ras) mutant; Receptor to advanced glycation end products (RAGE-1); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Receptor tyrosine protein kinase ERBB2 or Her-22 / neu; Renal pervasive protein 1 (RU1); Renal pervasive protein 2 (RU2); Sarcoma translocation breakpoint; Serine 2 (TMPRSS2) ETS fusion gene; Sialoyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialic acid Lewis antigen); Sperm protein 17 (SPA17); T cell recognized squamous cell carcinoma antigen 3 (SART3); Stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survival protein; Synovial sarcoma X breakpoint 2 (SSX2); TCR γ-substituted reading frame protein (TARP); TCR-β1 chain; TCR-β2 chain; TCR-δ chain; TCR-γ chain; TCRγ-δ; telomerase; TGFβR2; antigen recognized by TNT antibody;Thyroid-stimulating hormone receptor (TSHR); Timl- / HVCR1; Tissue factor 1 (TF1); Tn 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 associated (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-associated protein 2 (TRP-2); urolytic protein 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1); or X antigen family member 1A (XAGE1). In some embodiments, the targeting portion binds to CD7. In some embodiments, the targeting portion binds to CD8.

[0204] In some embodiments, the targeting portion binds to a target present on the cell. In some embodiments, the cells are selected from, but are 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, but not limited to, the group consisting of: 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.

[0205] CD7 binding polypeptide

[0206] In some implementations, the target moiety (e.g., a peptide) binds to CD7.

[0207] In some embodiments, the CD7-binding peptide is an antibody that binds to non-human primate CD7. In some embodiments, the CD7-binding peptide is an antibody that binds to human CD7. The sequence of human CD7 (UniProtKB P09564) is as follows (SEQ ID NO: 29):

[0208] MAGPPRLLLLPLLLALARGLPGALAAQEVQQSPHCTTVPVGASVNITCSTSGGLRGIYLRQLGPQPQDIIYYEDGVVPTTDRRFRGRIDFSGSQDNLTITMHRLQLSDTGTYTCQAITEVNV YGSGTLVLVTEEQSQGWHRCSDAPPRASALPAPPTGSALPDPQTASALPDPPAASALPAALAVISFLLGLGLGVACVLARTQIKKLCSWRDKNSAACVVYEDMSHSRCNTLSSPNQYQ (SEQ ID NO: 29).

[0209] In some embodiments, the peptide-bound CD7 is expressed on the cell surface. 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, α-β T cell, γ-δ T cell, lymphoprogenitor 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+.

[0210] 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 containing an Fc region. The Fc region may be linked to either the heavy or light chain of the antibody. In some embodiments, the Fc region is an IgG Fc. In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, the IgG Fc is an IgG1 Fc. In some embodiments, the antibody comprises a constant Fc region as shown herein, such as SEQ ID NO: 26, 27, or 28, or a mutant thereof as provided herein. In some embodiments, the identities of T, L1, Fc, L2, and X1 are as provided herein.

[0211] In some implementations, the target portion does not include the Fc region and is given by formula T-L3-X1. In some implementations, the identities of T, L3, and X1 are as provided herein.

[0212] In some embodiments, this document provides a peptide (e.g., a CD7-binding peptide). In some embodiments, this document provides an antibody (e.g., an anti-CD7 antibody). In some embodiments, the antibody is a recombinant antibody that binds to CD7. In some embodiments, the CD7 protein is the human CD7 protein. In some embodiments, the CD7 protein is a non-human CD7 protein (e.g., mouse, rat, pig, dog, non-human primate). As used herein, the term "recombinant antibody" refers to an antibody that is not naturally occurring. In some embodiments, the term "recombinant antibody" refers to an antibody that has not been isolated from a human subject.

[0213] In some embodiments, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antibody fragment comprises a peptide selected from the table below, which describes the CDR based on Kabat numbering.

[0214]

[0215] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to non-human primate CD7. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to human CD7. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 33-35. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 33. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 34. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 35. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having a sequence selected from SEQ ID NO: 30-32. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 30. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 31. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 32. The CDRs mentioned throughout the embodiments of this specification are interchangeable with CDRs characterized in different formats, such as Chothia and IMGT.

[0216] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof 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.

[0217] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, wherein 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.

[0218] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises: (i) a light chain having any of the aforementioned combinations of LCDR1, LCDR2, and LCDR3 sequences; and (ii) a heavy chain having any of the aforementioned combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0219] Different CDR motifs can be combined in any combination (including those not described in the table above). For example, the following embodiments are provided as non-limiting examples of such combinations.

[0220] 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 a variant of any of the foregoing.

[0221] Although the preceding paragraphs may refer to CDRs according to the Kabat system, equivalent CDR sequences from the names IMGT and CHOTHIA can be used.

[0222] 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.

[0223] In some embodiments, the polypeptide comprises a heavy chain variable region peptide or a variant thereof having one of the following sequences:

[0224] Table 2: CD7AB1 VH

[0225] SEQ ID NO: AB ID Number Sequence 36 CD7AB1 QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS

[0226] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences or a variant thereof:

[0227] Table 3: CD7AB1 VL

[0228] SEQ ID NO: AB ID Number Sequence 37 CD7AB1 DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR

[0229] In some embodiments, the peptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 36. H Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 37. L Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof contains V. H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36 or a variant thereof; and 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 comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36 or a variant thereof; and 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 comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36 or a variant thereof; and 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; and V L The peptide contains the sequence of SEQ ID NO: 37.

[0230] V H and V L The sequence can be of any form, including but not limited to the scFv form, where V H and V L The region connects to the peptide linker. Examples of peptide linkers that can be used to connect the various peptides described herein include, but are not limited to: (GGGGS) n(SEQ ID NO: 55), where each n is independently 1-5. 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 to the peptide linker. In some embodiments, the polypeptide comprises SEQ ID NO: 36 and SEQ ID NO: 37.

[0231] As provided in this article, peptides, antibodies, or their antigen-binding fragments can be sequence variants.

[0232] The sequences of peptides or antibodies can be modified to generate human IgG antibodies. Transformations of the sequences provided herein can be modified to generate other types of antibodies. CDRs can also be linked with other antibodies, proteins, or molecules to generate antibody fragments that bind to CD7.

[0233] In some embodiments, the peptides or antibodies provided herein are targeting portions on the surface of engineered viral particles. In some embodiments, the targeting portion allows binding to target cells. In some embodiments, the targeting portion is a CD7 binding portion, such as the peptides or antibodies provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38.

[0234] DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGG GSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS (SEQ ID NO: 38)

[0235] Alternatively, it may be substantially similar to SEQ ID NO: 38, or an active fragment of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises the sequence shown in SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) shown 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.

[0236] In some embodiments, the peptide or antibody, as provided herein, is a targeting portion 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 portion allows binding to target cells. In some embodiments, the targeting portion is a CD7 binding portion, such as the peptide or antibody provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) comprises at least 90% of the sequence of SEQ ID NO: 39.

[0237] QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGS GGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (SEQ ID NO: 39)

[0238] Alternatively, it may be substantially similar to SEQ ID NO: 39, or an active fragment of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) as shown 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.

[0239] CD8 binding polypeptide

[0240] In some implementations, the targeting portion (peptide) can bind to CD8.

[0241] In some embodiments, the peptide binds to CD8. In some embodiments, the peptide binds to CD8-α. In some embodiments, the peptide binds to CD8-β. In some embodiments, the peptide binds to a CD8 heterodimer. In some embodiments, the CD8 heterodimer comprises CD8-α and CD8-β subunits. In some embodiments, the peptide binds to a CD8-α homodimer. In some embodiments, the peptide binding to CD8 is an antibody binding to non-human primate CD8. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8-α. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8-β. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8-α homodimer. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8 heterodimer. In some embodiments, the CD8-binding peptide 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-α. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-β. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-α 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-α (UniProtKB Q8TAW8) is as follows (SEQ ID NO: 40):

[0242] MALPPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGCYFCSALS NSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV (SEQ ID NO: 40).

[0243] The sequence of human CD8-β (UniProtKB Q8TD28) is as follows (SEQ ID NO: 41):

[0244] MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKISLSNMRIYWLRQRQAPSSDSHHEFLALWDSAKGTIHGEEVEQEKIAVFRDASRFILNLTSVK PEDSGIYFCMIVGSPELTFGKGTQLSVVDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPITLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQLYK (SEQ ID NO:41).

[0245] In some embodiments, the peptide-bound CD8 is expressed on the cell surface. 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, α-β T cell, γ-δ T cell, lymphoprogenitor 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.

[0246] 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 containing an Fc region. The Fc region may be linked to either the heavy or light chain of the antibody. In some embodiments, the Fc region is an IgG Fc. In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, the IgG Fc is an IgG1 Fc. In some embodiments, the antibody comprises a constant Fc region as shown herein, such as SEQ ID NO: 26, 27, or 28, or variants thereof as provided herein. In some embodiments, the identities of T, L1, Fc, L2, and X1 are as provided herein.

[0247] In some implementations, the target portion does not include the Fc region and is given by formula T-L3-X1. In some implementations, the identities of T, L3, and X1 are as provided herein.

[0248] In some embodiments, this document provides a peptide (e.g., a CD8-binding peptide). In some embodiments, this document provides an antibody (e.g., an anti-CD8 antibody). 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, non-human primate). As used herein, the term "recombinant antibody" refers to an antibody that is not naturally occurring. In some embodiments, the term "recombinant antibody" refers to an antibody that has not been isolated from a human subject.

[0249] In some embodiments, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antibody fragment comprises a peptide selected from the table below, which describes the CDR based on Kabat numbering.

[0250]

[0251] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to non-human primate CD8. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to human CD8. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 45-47. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 45. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 46. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 47. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having a sequence selected from SEQ ID NO: 42-44. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 42. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 43. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 44. The CDRs mentioned throughout the embodiments of this specification may be interchanged with CDRs characterized in different formats, such as Chothia and IMGT.

[0252] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof 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.

[0253] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, wherein 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.

[0254] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises: (i) a light chain having any of the aforementioned combinations of LCDR1, LCDR2, and LCDR3 sequences; and (ii) a heavy chain having any of the aforementioned combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0255] Different CDR motifs can be combined in any combination (including those not described in the table above). For example, the following embodiments are provided as non-limiting examples of such combinations.

[0256] 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 a variant of any of the foregoing.

[0257] Although the preceding paragraphs may refer to CDRs according to the Kabat system, equivalent CDR sequences from the names IMGT and CHOTHIA can be used.

[0258] 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.

[0259] In some embodiments, the polypeptide comprises a heavy chain variable region peptide or a variant thereof having one of the following sequences:

[0260] Table 5: CD8AB1 VH

[0261] SEQ ID NO: AB ID Number Sequence 48 CD8AB1 QVQLVQSGAEVKKPGASVKISCKASGYTFTSYWISWVRQAPGQGLEWMGDIYPDSNTGSGTYLYSLQSISRDNSKNTLYLQMNSLRAEDTAVYYCAADVSSGTYFDYWGQGTLVTVSS EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS

[0262] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences or a variant thereof:

[0263] Table 6: CD8AB1 VL

[0264] SEQ ID NO: AB ID Sequence 49 CD8AB1 NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR

[0265] In some embodiments, the peptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 48. H Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 49. L Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof contains V. H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 48 or a variant thereof; and 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 comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 48 or a variant thereof; and 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 comprises V H Peptides and Vitamins L peptides, of which VH The peptide contains the sequence of SEQ ID NO: 48 or a variant thereof; and 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; and V L The peptide contains the sequence of SEQ ID NO: 49.

[0266] V H and V L The sequence can be of any form, including but not limited to the scFv form, where V H and V L The region connects to the peptide linker. Examples of peptide linkers that can be used to connect the various peptides described herein include, but are not limited to: (GGGGS) n (SEQ ID NO: 55), where each n is independently 1-5. 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 to the peptide linker. In some embodiments, the polypeptide comprises SEQ ID NO: 48 and SEQ ID NO: 49.

[0267] The sequences of peptides or antibodies can be modified to generate human IgG antibodies. Transformations of the sequences provided herein can be modified to generate other types of antibodies. CDRs can also be linked with other antibodies, proteins, or molecules to generate antibody fragments that bind to CD8.

[0268] In some embodiments, the peptides or antibodies provided herein are targeting portions on the surface of engineered viral particles. In some embodiments, the targeting portion allows binding to target cells. In some embodiments, the target-binding domain (“T”) is a CD8 binding portion, such as the peptides or antibodies provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 50.

[0269] NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSG GGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS (SEQ ID NO: 50)

[0270] 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”) comprises at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises the sequence shown in SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) as shown 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.

[0271] In some embodiments, the peptide or antibody, as provided herein, is a targeting portion 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 portion allows binding to target cells. In some embodiments, the target-binding domain (“T”) is a CD8 binding portion, such as the peptide or antibody provided herein. In some embodiments, the target-binding domain (“T”) comprises 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.

[0272] EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGG GGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (SEQ ID NO: 51)

[0273] Alternatively, it may be substantially similar to SEQ ID NO: 51, or an active fragment of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises the sequence shown in SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) shown 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.

[0274] Targeting moiety comprising an Fc domain

[0275] In some implementation schemes, V H and V LThe polypeptide is linked to a stem portion S1 containing the Fc region. In some embodiments, the Fc region is as provided herein. In some embodiments, the Fc region is a variant Fc region as provided herein. Non-restrictive mutations in the Fc region are provided herein. In some embodiments, the mutant Fc region contains a sequence as a variant of SEQ ID NO:26, SEQ ID NO:27, or SEQ ID NO:28 as provided herein. In some embodiments, the variant of SEQ ID NO:26 contains one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A as provided herein. In some embodiments, the variant of SEQ ID NO:27 contains one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A as 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 as provided herein. As provided herein, the heavy chain may be linked to the Fc region. In some embodiments, the Fc region also comprises (e.g., linked to) a transmembrane domain. In some embodiments, the Fc region also comprising the transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising the transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. Examples of ECDs include, but are not limited to, the CD8 and / or CD28 extracellular domains as described herein. T M Examples include, but are not limited to, CD8 and / or CD28 transmembrane domains as provided herein. In some embodiments, X1 includes T M Furthermore, ECD and ICD are not present. In some implementations, X1 includes ECD and T. M And ICD does not exist. In some implementations, X1 includes T. M And ICD, but ECD is not present. In some implementations, X1 includes ECD, T MAnd ICD. In any of the following embodiments, it should be understood that ECD, ICD, or both may optionally be absent. Thus, X1 comprises CD8 and / or CD28 ECD, CD8 and / or CD28 T. M Implementations of ICDs containing Env-doped motifs are understood to include the following X1 members: i) CD8 and / or CD28 ECD, CD8 and / or CD28 T M and ICDs containing Env-doped motifs; ii) CD8 and / or CD28 T M and ICDs containing Env-doped motifs, where ECDs are absent; iii) CD8 and / or CD28 ECDs and CD8 and / or CD28 T M iv) where ICD is absent; and iv) CD8 and / or CD28 T M Where neither ECD nor ICD exists. Similarly, where X1 contains CD8 and / or CD28 T. M Implementations of ICDs containing Env-doped motifs are understood to include the following X1 members: i) CD8 and / or CD28 T M and ICDs containing Env-doped motifs; and ii) CD8 and / or CD28 T M Where ICD is absent. Similarly, where X1 contains CD8 and / or CD28 ECD and CD8 and / or CD28 T. M The implementation scheme is understood to include the following X1 members: i) CD8 and / or CD28 ECD and CD8 and / or CD28 T M ; and ii) CD8 and / or CD28 T M The ECD is not present. It should also be understood that the foregoing explanation does not address specific ECDs or Ts. M This is also correct in the implementation scheme of the ICD. For example, where X1 includes an ECD, CD8, and / or CD28 T. M The implementation scheme of ICD will be understood to include the following X1 members: i) ECD, CD8 and / or CD28 T M and ICD; ii) ECD and CD8 and / or CD28 T M iii) CD8 and / or CD28 T M and ICD, where ECD is absent; and iv) CD8 and / or CD28 T M Where ECD and ICD are absent. Similarly, where X1 contains CD8 and / or CD28 T. M The implementation scheme of ICD is understood to include the following X1 members: i) CD8 and / or CD28T Mand ICD; and ii) CD8 and / or CD28 T M Where ICD is absent. Similarly, where X1 contains ECD and CD8 and / or CD28T. M The implementation scheme is understood to include the following X1 members: i) ECD and CD8 and / or CD28 T M ; and ii) CD8 and / or CD28T M Where the ECD does not exist. Unless otherwise stated, the foregoing examples and explanations apply to any of the following implementations.

[0276] In some implementations, X1 includes CD8 and / or CD28 ECD, T M And ICD. In some implementations, X1 includes an ECD, CD8, and / or CD28 T. M and ICD. In some implementations, X1 includes CD8 and / or CD28 ECD, CD8 and / or CD28 TCD. M and ICD. In some implementations, X1 includes ECD, T M and ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, X1 contains CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, X1 comprises ECD, CD8, and / or CD28 T. M And ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, X1 comprises CD8 and / or CD28 ECD, CD8 and / or CD28 TCD. M and ICD, wherein the ICD contains env-incorporated motifs as provided herein.

[0277] In some implementation schemes, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing the Fc region as described herein. In some embodiments, the V region described herein... H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region comprising a transmembrane domain as described herein. In some embodiments, the Fc region comprising the transmembrane domain also has the formula L1-Fc-L2-X1, where L1 is a linker as described herein or is absent, Fc is a variant Fc region as described herein, L2 is a linker as described herein or is absent, and X1 is a polypeptide comprising the transmembrane domain as described herein. As provided herein, X1 may comprise a portion having the formula ECD-T M-ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or does not exist.

[0278] In some implementation schemes, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M and ICD. In some implementations, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M and ICD. In some implementations, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M and ICD. In some implementations, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M and ICD, wherein the ICD contains an Env-incorporated motif as provided herein. In some embodiments, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M and ICD, wherein the ICD contains an Env-incorporated motif as provided herein. In some embodiments, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M and ICD, wherein the ICD contains an Env-incorporated motif as provided herein. In some embodiments, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. Mand ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the V provided herein is linked to the stem portion (S1) (L1-Fc-L2-X1) containing the Fc region. H and V L The peptide anchors to the surface of viral particles (such as those described in this article), and the Fc region contains ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0279] In some implementations, V having a sequence such as SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M and ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H peptides and V having the sequence shown in SEQ ID NO:37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M and ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M and ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 LThe peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 36. H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 36. H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 36. H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V with a sequence as shown in SEQ ID NO:36 is attached to the stem portion (S1) (L1-Fc-L2-X1) containing the Fc region. H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide anchors to the surface of viral particles (such as those described in this article), and the Fc region contains ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0280] In some embodiments, V 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: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:37. L The peptide is linked to a stem portion (S1) comprising the Fc region as described herein. In some embodiments, a V-shaped portion comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide is linked to a stem portion (S1) containing an Fc region comprising a transmembrane domain as described herein. In some embodiments, the Fc region containing the transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker as described herein or is absent, Fc is a variant Fc region as described herein, L2 is a linker as described herein or is absent, and X1 is a polypeptide containing the transmembrane domain as described herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, V 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: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. MAnd ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M And ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28T. M And ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. HPeptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M And ICD, wherein the ICD contains the Env incorporation motif as provided herein. In some embodiments, V 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: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V 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: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V 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: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37.L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the V, which contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36, is linked to the stem portion (S1) (L1-Fc-L2-X1) containing the Fc region. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide anchors to the surface of viral particles (such as those described in this article), and the Fc region contains ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0281] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37.

[0282] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V LThe peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V H Peptides and Vitamins L The peptide comprises a light chain CDR having the sequence of SEQ ID NO: 33-35; and / or a heavy chain CDR having the sequence of SEQ ID NO: 30-32. In some embodiments, the polypeptide comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V H Peptides and Vitamins 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 shown in the combinations provided in this article.

[0283] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V LThe 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; and V H The peptide comprises 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.

[0284] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 33, wherein LCDR1 contains at most one conserved amino acid substitution; LCDR2 having the sequence of SEQ ID NO: 34, wherein LCDR2 contains at most one conserved amino acid substitution; and LCDR3 having the sequence of SEQ ID NO: 35, wherein LCDR3 contains at most one conserved amino acid substitution; and V H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 30, wherein HCDR1 contains at most one conserved amino acid substitution; HCDR2 having the sequence of SEQ ID NO: 31, wherein HCDR2 contains at most one conserved amino acid substitution; and HCDR3 having the sequence of SEQ ID NO: 32, wherein HCDR3 contains at most one conserved amino acid substitution.

[0285] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36, and V L The peptide contains the sequence of SEQ ID NO: 37.

[0286] In some embodiments, the peptides provided herein bind to non-human primate CD7. In some embodiments, the peptides provided herein bind to human CD7.

[0287] As provided in this article, the different polypeptides (V) described herein H or V L This can be linked to a peptide linker or not to form a continuous sequence. In some embodiments, the peptide linker comprises (GGGGS). n The sequence (SEQ ID NO: 55), wherein each n is independently 1-5. 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 can be derived from formula V. H -ZV L or V L -ZV H This indicates that Z represents a peptide linker. In some embodiments, Z is (GGGGS). n (SEQ ID NO: 55), where each n is independently 1-5. 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.

[0288] In some implementations, it includes formula V L -ZV H The peptide representing the linked peptide comprises a heavy chain variable region, as shown in SEQ ID NO: 36, linked to a light chain variable region, as shown in SEQ ID NO: 37, via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, it comprises a heavy chain variable region, as shown in SEQ ID NO: 36, linked to a light chain variable region, as shown in SEQ ID NO: 37, via a peptide linker. H Connected V L The polypeptide has the sequence shown below.

[0289] DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGG GSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS (SEQ ID NO: 38).

[0290] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises at least 90% of the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises at least 95% of the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises at least 99% of the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises the sequence shown in SEQ ID NO: 38. In some embodiments, the polypeptide shown in SEQ ID NO: 38 is an antibody or an antigen-binding fragment thereof. In some embodiments, the antibody is an anti-CD7 antibody.

[0291] In some implementations, it includes formula V H -ZV L The peptide representing the linked peptide comprises a light chain variable region, as shown in SEQ ID NO: 37, linked to a heavy chain variable region, as shown in SEQ ID NO: 36, via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, it comprises a light chain variable region, as shown in SEQ ID NO: 37, linked to a heavy chain variable region, as shown in SEQ ID NO: 36, via a peptide linker. L Connected V H The polypeptide has the sequence shown below.

[0292] QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGS GGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (SEQ ID NO: 39).

[0293] In some embodiments, the polypeptide comprises 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 comprises at least 90% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises the sequence shown in SEQ ID NO: 39. In some embodiments, the polypeptide shown in SEQ ID NO: 39 is an antibody or an antigen-binding fragment thereof. 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.

[0294] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region, which further comprises transmembrane domains, such as those provided herein. In some embodiments, the Fc region comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T MIt is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes an ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, the Fc region comprising ECD, T MAnd ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains the Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising 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 comprising a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising an ECD, T M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0295] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region, which further comprises transmembrane domains, such as those provided herein. In some embodiments, the Fc region comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes an ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. MAnd ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, the Fc region comprising ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T MAnd ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising 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 comprising a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising an ECD, T M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0296] In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and a stem portion (S1) containing an Fc region, said Fc region further comprising a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L. 1- Fc-L2-X1, where L1 is a linker as described herein or is absent, Fc is a variant Fc region as described herein, L2 is a linker as described herein or is absent, and X1 is a polypeptide containing a transmembrane domain as described herein. As described herein, X1 may contain a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T MAnd ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO:38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising a sequence having the sequence shown in SEQ ID NO: 38 and containing a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region containing an ECD, T M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0297] In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and a stem portion (S1) containing an Fc region, the Fc region further comprising a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a polypeptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO:39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T MAnd ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M and ICD, wherein the ICD contains the Env doped motif as provided herein.

[0298] In some embodiments, a polypeptide comprising a sequence having the sequence shown in SEQ ID NO: 39 and including a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0299] In some embodiments, the polypeptide comprising formula T-S1 (where T is a target-binding domain and S1 is a stem portion) provided herein comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the amino acid sequence identical to that of SEQ ID NO: 83.

[0300] (SEQ ID NO: 83)

[0301] Alternatively, it may be substantially similar to SEQ ID NO: 83, or an active fragment of SEQ ID NO: 83. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 90% of the amino acid sequence identical to that of SEQ ID NO: 83. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 95% of the amino acid sequence identical to that of SEQ ID NO: 83. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 98% of the amino acid sequence identical to that of SEQ ID NO: 83. In some embodiments, the polypeptide comprising T-S1 provided herein contains the amino acid sequence of SEQ ID NO: 83.

[0302] In some embodiments, the polypeptide comprising formula T-S1 provided herein (where T is a target-binding domain and S1 is a stem portion) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of 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 wherein the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises at least 90% of the amino acid 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 wherein the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein 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, and wherein the stem portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein 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, and wherein the stem portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises the amino acid 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 wherein the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83.

[0303] In some embodiments, the polypeptide of formula T-S1 provided herein (where T is a polypeptide containing V) H and V L The target-binding domain (and S1 is the stem portion) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain HThe V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L The amino acids 172-283 correspond to those of SEQ ID NO: 83, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L The amino acids 172-283 correspond to those in SEQ ID NO: 83, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein contains at least 95% of the same amino acid sequence as the sequence in SEQ ID NO: 83, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L The amino acids 172-283 correspond to those of SEQ ID NO: 83, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L The amino acids 172-283 of SEQ ID NO: 83 correspond to amino acids 284-634 of SEQ ID NO: 83, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises the amino acid sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L The amino acids 172-283 of SEQ ID NO: 83 correspond to amino acids 284-634 of SEQ ID NO: 83, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 83.

[0304] In some embodiments, the polypeptide of the formula T-L1-Fc-L2-X1 provided herein (where T is a target-binding domain, L1 is a polypeptide linker or is absent, Fc is a variant Fc domain as provided herein, L2 is a polypeptide linker or is absent, and X1 is a polypeptide containing a transmembrane domain) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of 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 of formula T-L1-Fc-L2-X1 provided herein contains at least 90% of the same amino acid sequence as 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, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, the polypeptide of formula T-L1-Fc-L2-X1 provided herein contains at least 95% of the same amino acid sequence as 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, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83. In some embodiments, the polypeptide of formula T-L1-Fc-L2-X1 provided herein contains at least 98% of the same amino acid sequence as 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, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83.In some embodiments, the polypeptide of formula T-L1-Fc-L2-X1 provided herein contains the amino acid 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, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 83.

[0305] In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein (where T is a polypeptide comprising V) H and V L The target-binding domain, L1 is a peptide linker or is absent, Fc is a variant Fc domain as provided herein, L2 is a peptide linker or is absent, and X1 is a peptide containing a transmembrane domain) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequence to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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 comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 90% of the amino acid sequence identical to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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 comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 95% of the same amino acid sequence as the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain HThe V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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 comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 98% identical amino acid sequence to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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 comprising the formula T-L1-Fc-L2-X1 provided herein contains the amino acid sequence of SEQ ID NO: 83, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L The amino acids 172-283 of SEQ ID NO: 83 correspond to L1, which contains amino acids 284-301 of SEQ ID NO: 83, and Fc which 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.

[0306] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD peptide (where T is the target-binding domain, L1 is the peptide linker or absent, Fc is the variant Fc domain as provided in this paper, L2 is the peptide linker or absent, ECD is the extracellular domain, T M(It is a transmembrane domain, and the ICD is an intracellular domain containing an env-incorporated motif) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences 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 ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains at least 90% identical amino acid sequence to that 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 ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains at least 95% identical amino acid sequence to that 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... MThe ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains at least 98% identical amino acid sequence to that 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 ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of -ICD contains the amino acid 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 env-incorporated motif comprises amino acids 585-612 of SEQ ID NO: 83, and the ICD comprises amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif comprises amino acids 627-634 of SEQ ID NO: 83.

[0307] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD polypeptide (where T is the peptide containing V) H and V L The target-binding domain, L1 is a peptide linker or is absent, Fc is the variant Fc domain as provided in this paper, L2 is a peptide linker or is absent, ECD is an extracellular domain, T M(It is a transmembrane domain, and the ICD is an intracellular domain containing an env-incorporated motif) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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, and T M The ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 90% of the same amino acid sequence as SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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, and T M The ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 95% identical amino acid sequence to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. LCorresponding 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, and T M The ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 98% identical amino acid sequence to the sequence of SEQ ID NO:83, wherein the V target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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, and T M The ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains the amino acids of SEQ ID NO: 83, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 83. L Corresponding 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, and T M The ICD contains amino acids 585-612 of SEQ ID NO: 83, and the ICD contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 83.

[0308] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD peptide (where T is the target-binding domain, L1 is the peptide linker or absent, Fc is the variant Fc domain as provided in this paper, L2 is the peptide linker or absent, ECD is the extracellular domain, T M The transmembrane domain, and the ICD (an intracellular domain containing an env-incorporated motif), contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence 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, and T... M The ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains 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 contains the amino acid sequence 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 MThe ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains 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 contains the amino acid sequence 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 ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains 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 contains the amino acid sequence 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 MThe ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains the amino acid sequence of SEQ ID NO: 83, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence 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 comprising SEQ ID NO: 62 and corresponding to amino acids 585-612 of SEQ ID NO: 83, and the ICD comprising amino acids 613-634 of SEQ ID NO: 83, wherein the env incorporation motif comprises the amino acid sequence comprising SEQ ID NO: 63 and corresponding to amino acids 627-634 of SEQ ID NO: 83.

[0309] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD polypeptide (where T is the peptide containing V) H and V L The target-binding domain, L1 is a peptide linker or is absent, Fc is the variant Fc domain as provided in this paper, L2 is a peptide linker or is absent, ECD is an extracellular domain, T M (It is a transmembrane domain, and the ICD is an intracellular domain containing an env-incorporated motif) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 83, V of the target-binding domain.L The sequence 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 ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 90% of the same amino acid sequence as SEQ ID NO: 83, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 83, V of the target-binding domain. L The sequence 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 ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... MThe -ICD polypeptide contains at least 95% of the same amino acid sequence as SEQ ID NO: 83, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 83, V of the target-binding domain. L The sequence 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 ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 98% identical amino acid sequence to the sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 83, V of the target-binding domain. L The sequence 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 MThe ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 83, and contains amino acids 613-634 of SEQ ID NO: 83, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 83. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains the amino acid sequence of SEQ ID NO: 83, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 83, V of the target-binding domain. L The sequence 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 comprising SEQ ID NO: 62 and corresponding to amino acids 585-612 of SEQ ID NO: 83, and the ICD comprising amino acids 613-634 of SEQ ID NO: 83, wherein the env incorporation motif comprises the amino acid sequence comprising SEQ ID NO: 63 and corresponding to amino acids 627-634 of SEQ ID NO: 83.

[0310] In some implementations, V having a sequence such as SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H peptides and V having the sequence shown in SEQ ID NO:49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T MAnd ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 48. H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 48. H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M and ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 48. H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. Mand ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V with a sequence as shown in SEQ ID NO:48 is attached to the stem portion (S1) (L1-Fc-L2-X1) containing the Fc region. H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide anchors to the surface of viral particles (such as those described in this article), and the Fc region contains ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0311] In some embodiments, V 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: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:49. L The peptide is linked to a stem portion (S1) comprising the Fc region as described herein. In some embodiments, a V-shaped portion comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) containing an Fc region comprising a transmembrane domain as described herein. In some embodiments, the Fc region containing the transmembrane domain has the formula L. — Fc-L2-X1, where L1 is a linker as described herein or is absent, Fc is a variant Fc region as described herein, L2 is a linker as described herein or is absent, and X1 is a polypeptide containing a transmembrane domain as described herein. As described herein, X1 may contain a peptide having the formula ECD-T M-ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, V 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: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M And ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M And ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28T. MAnd ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing the Fc region, which includes ECD and T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V 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: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes CD8 and / or CD28 ECD, T M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V 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: 48.H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a V 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: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the V, which contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 48, is linked to the stem portion (S1) (L1-Fc-L2-X1) containing the Fc region. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide anchors to the surface of viral particles (such as those described in this article), and the Fc region contains ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0312] In some implementations, the polypeptide contains V H Peptides and Vitamins Lpeptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 48; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49.

[0313] In some implementations, V is provided H Peptides and Vitamins L peptides are polypeptides, in which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO:48; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 49; the condition is V H Peptides and Vitamins L The peptide comprises a light chain CDR having the sequence of SEQ ID NO: 45-47; and / or a heavy chain CDR having the sequence of SEQ ID NO: 42-44. In some embodiments, the polypeptide comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 48; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO:49; the condition is V. H Peptides and Vitamins LThe 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 shown in the combinations provided in this article.

[0314] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 48; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 49; the condition is 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; and V H The peptide comprises 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.

[0315] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 48; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 49; the condition is V LThe peptide comprises LCDR1 having the sequence of SEQ ID NO: 45, wherein LCDR1 contains at most one conserved amino acid substitution; LCDR2 having the sequence of SEQ ID NO: 46, wherein LCDR2 contains at most one conserved amino acid substitution; and LCDR3 having the sequence of SEQ ID NO: 47, wherein LCDR3 contains at most one conserved amino acid substitution; and V H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 42, wherein HCDR1 contains at most one conserved amino acid substitution; HCDR2 having the sequence of SEQ ID NO: 43, wherein HCDR2 contains at most one conserved amino acid substitution; and HCDR3 having the sequence of SEQ ID NO: 44, wherein HCDR3 contains at most one conserved amino acid substitution.

[0316] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 48, and V L The peptide contains the sequence of SEQ ID NO: 49.

[0317] In some embodiments, the peptides provided herein bind to non-human primate CD8. In some embodiments, the peptides provided herein bind to human CD8.

[0318] As provided in this article, the different polypeptides (V) described herein H or V L This can be linked to a peptide linker or not to form a continuous sequence. In some embodiments, the peptide linker comprises (GGGGS). n The sequence (SEQ ID NO: 55), wherein each n is independently 1-5. 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 can be derived from formula V. H -ZV L or V L -ZV H This indicates that Z represents a peptide linker. In some embodiments, Z is (GGGGS). n (SEQ ID NO: 55), where each n is independently 1-5. 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.

[0319] In some implementations, it includes formula V L -ZV H The peptide representing the linked peptide comprises a heavy chain variable region, as shown in SEQ ID NO: 48, linked to a light chain variable region, as shown in SEQ ID NO: 49, via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, it comprises a heavy chain variable region, as shown in SEQ ID NO: 48, linked to a light chain variable region, as shown in SEQ ID NO: 49, via a peptide linker. H Connected V L The polypeptide has the following sequence:

[0320] NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSG GGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS (SEQ ID NO: 50).

[0321] In some embodiments, the polypeptide comprises 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 comprises at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide comprises at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide comprises at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide comprises the sequence shown in SEQ ID NO: 50. In some embodiments, the polypeptide shown in SEQ ID NO: 50 is an antibody or an antigen-binding fragment thereof. In some embodiments, the antibody is an anti-CD8 antibody.

[0322] In some implementations, it includes formula V H -ZV LThe peptide representing the linked peptide comprises a light chain variable region, as shown in SEQ ID NO: 49, linked to a heavy chain variable region, as shown in SEQ ID NO: 48, via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, it comprises a light chain variable region, as shown in SEQ ID NO: 49, linked to a heavy chain variable region, as shown in SEQ ID NO: 48, via a peptide linker. L Connected V H The polypeptide has the following sequence:

[0323] EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGG GGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (SEQ ID NO: 51).

[0324] In some embodiments, the polypeptide comprises 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 comprises at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide comprises at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide comprises at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide comprises the sequence shown in SEQ ID NO: 51. In some embodiments, the polypeptide shown in SEQ ID NO: 51 is an antibody or an antigen-binding fragment thereof. 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.

[0325] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 50 and comprises a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 50 and comprises a stem portion (S1) containing an Fc region, which further comprises transmembrane domains, such as those provided herein. In some embodiments, the Fc region comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes an ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. MAnd ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, the Fc region comprising ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains the Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 50 and comprises a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T MAnd ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising 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 comprising a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising an ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0326] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 51 and comprises a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 51 and comprises a stem portion (S1) containing an Fc region, which further comprises transmembrane domains, such as those provided herein. In some embodiments, the Fc region comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T MAnd ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, the Fc region comprising ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T MAnd an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises 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 stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising 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 comprising a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising an ECD, T M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0327] In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and a stem portion (S1) containing an Fc region, said Fc region further comprising a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide containing a transmembrane domain as provided herein. As provided herein, X1 may comprise a polypeptide having the formula ECD-TM -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO:50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, the Fc region containing ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T MAnd ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 50 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising a sequence having the sequence shown in SEQ ID NO: 50 and containing a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region containing an ECD, T M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0328] In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and a stem portion (S1) containing an Fc region, the Fc region further comprising a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a polypeptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, which includes ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T MAnd ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO:51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region, the Fc region containing ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 51 and includes a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an Env incorporation motif as provided herein. In some embodiments, a polypeptide comprising a sequence having the sequence shown in SEQ ID NO:51 and containing a stem portion (S1) (L1-Fc-L2-X1) containing an Fc region is anchored to the surface of a viral particle (such as those provided herein), the Fc region containing an ECD, T... M And ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0329] Targeting moiety comprising a flexible polypeptide

[0330] In some implementation schemes, V H and V L The polypeptide is linked to a stem portion (S1) comprising a flexible polypeptide (L3) as provided herein. In some embodiments, V H and V L The polypeptide is linked to a stem portion (S1) comprising a flexible polypeptide, which further comprises a polypeptide containing a transmembrane domain. In some embodiments, the flexible polypeptide comprising a polypeptide containing a transmembrane domain is represented by the formula L3-X1, wherein L3 is a flexible polypeptide as provided herein, and X1 is as provided herein. As provided herein, X1 may comprise a polypeptide having the formula ECD-T M -ICD polypeptides, wherein the ECD is an extracellular domain or a fragment thereof as provided herein, or is absent; T M It is a transmembrane domain as described herein; and the ICD is an intracellular domain as described herein or is absent. Examples of ECDs include, but are not limited to, CD8 and / or CD28 ECDs as described herein. M Examples include, but are not limited to, CD8 and / or CD28 transmembrane domains as provided herein. In any of the following embodiments, it should be understood that ECD, ICD, or both may optionally be absent. Thus, X1 comprises CD8 and / or CD28 ECD, CD8 and / or CD28 T... M Implementations of ICDs containing Env-doped motifs are understood to include the following X1 members: i) CD8 and / or CD28 ECD, CD8 and / or CD28 T M and ICDs containing Env-doped motifs; ii) CD8 and / or CD28 T M and ICDs containing Env-doped motifs, where ECDs are absent; iii) CD8 and / or CD28 ECDs and CD8 and / or CD28 Ts M iv) where ICD is absent; and iv) CD8 and / or CD28 T M Where neither ECD nor ICD exists. Similarly, where X1 contains CD8 and / or CD28 T. M Implementations of ICDs containing Env-doped motifs are understood to include the following X1 members: i) CD8 and / or CD28T M and ICDs containing Env-doped motifs; and ii) CD8 and / or CD28 T M Where ICD is absent. Similarly, where X1 contains CD8 and / or CD28 ECD and CD8 and / or CD28 T. MThe implementation scheme is understood to include the following X1 members: i) CD8 and / or CD28 ECD and CD8 and / or CD28 T M ; and ii) CD8 and / or CD28 T M The ECD is not present. It should also be understood that the foregoing explanation does not address specific ECDs or Ts. M This is also correct in the implementation scheme of the ICD. For example, where X1 includes an ECD, CD8, and / or CD28 T. M The implementation of ICD will be understood to include the following X1 members: i) ECD, CD8 and / or CD28 T M and ICD; ii) ECD and CD8 and / or CD28 T M iii) CD8 and / or CD28 T M and ICD, where ECD is absent; and iv) CD8 and / or CD28 T M Where ECD and ICD are absent. Similarly, where X1 contains CD8 and / or CD28 T. M The implementation scheme of ICD is understood to include the following X1 members: i) CD8 and / or CD28 T M and ICD; and ii) CD8 and / or CD28 T M Where ICD is absent. Similarly, where X1 contains ECD and CD8 and / or CD28 T. M The implementation scheme is understood to include the following X1 members: i) ECD and CD8 and / or CD28 T M ; and ii) CD8 and / or CD28 T M Where the ECD does not exist. Unless otherwise stated, the foregoing examples and explanations apply to any of ...

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 comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety.

2. The viral particle of claim 1, wherein the gene editing system is a CRISPR-Cas system, a zinc finger nuclease system, a TALEN, a meganuclease, or a gene product modulating nucleic acid molecule.

3. The viral particle of claim 2, wherein 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 a target nucleic acid molecule.

4. The viral particle of claim 2, wherein the CRISPR-Cas system is a Class 1 or Class 2 CRISPR-Cas system.

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

6. The viral particle of claim 5, wherein the Type II Cas protein is a Cas9 protein.

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

8. The viral particle of claim 7, wherein the Type V Cas protein is selected from Casl2a, Casl2b, Casl2c, Casl2d, Casl2e, or Cas 14.

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

10. The viral particle of claim 9, wherein the Type VI Cas protein is selected from Casl3a, Casl3b, Casl3c, or Casl3d.

11. The viral particle of claim 2, wherein the CRISPR-Cas system is a base editing system.

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

13. The viral particle of claim 2, wherein the CRISPR-Cas system is a prime editing system.

14. The viral particle of claim 2, wherein the CRISPR-Cas system comprises a CRISPRa, a CRISPRi, a Cas 3, or a CasMINI protein.

15. The viral particle of claim 2, wherein the gene product modulating nucleic acid molecule encodes at least one molecule selected from an siRNA, a piRNA, a miRNA, an RNAi RNA, an mRNA, an shRNA, and an antisense RNA.

16. The viral particle of claim 1, wherein the stalk moiety S1 comprises a variant Fc protein, wherein the variant Fc protein comprises a transmembrane domain, and wherein the mutant Fc protein comprises at least one effector mutation, wherein the effector mutation inhibits interaction between the Fc protein and an Fc interacting protein such as FcyR, C1q, FcRb, or FcRn.

17. The viral particle of claim 16, wherein the S1 stalk moiety is attached to the surface of the viral particle via the transmembrane domain.

18. The viral particle of claim 16, wherein the Fc protein is an IgG1 Fc, IgG2 Fc, or IgG4 Fc protein.

19. The viral particle of 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 viral particle of claim 16, wherein the variant Fc protein is a variant IgG1 Fc protein.

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

22. The viral particle of claim 20, wherein the variant IgG1 Fc protein comprises an amino acid sequence that is at least 80% identical to SEQ ID NO: 82, at least 85% identical to SEQ ID NO: 82, at least 90% identical to SEQ ID NO: 82, at least 95% identical to SEQ ID NO: 82, at least 98% identical to SEQ ID NO: 82, or at least 100% identical to SEQ ID NO:

82.

23. The viral particle of claim 1, wherein the targeting moiety having the formula T-S1 comprises a stalk moiety S1 having the formula L1-Fc-L2-X1, wherein: L1 is a linker or is absent; Fc is a variant Fc protein; L2 is a linker or is absent; and X1 is a polypeptide comprising a transmembrane domain, wherein the targeting moiety having the formula T-S1 has the formula T-L1-Fc-L2-X1.

24. The viral particle of claim 23, wherein L1 and L2 are each independently absent or a polypeptide linker comprising 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. ECD is an extracellular domain of a cell surface protein or fragment thereof, or is absent; ​ ​ ​ ​ 25. The viral particle of claim 24, wherein X1comprises a polypeptide having the formula ECD-T M -ICD, wherein: ​ T M is a transmembrane domain of a transmembrane protein; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, or is absent, wherein said targeting moiety having said formula T-L1-Fc-L2-X1 has the formula T-L1-Fc-L2-ECD-T M - ICD.

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

60.

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

62.

28. The viral particle of claim 25, wherein the ICD comprises an Env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO:

64.

29. The viral particle of claim 1, wherein the target-binding domain (T) binds to: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR α, TCR β, TCR γ, TCR δ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7 or CXCR3, glycosylated CD43 epitopes expressed on acute leukemia or lymphoma but not on hematopoietic progenitor cells, glycosylated CD43 epitopes expressed on non-hematopoietic cancers, A kinase anchoring protein 4 (AKAP-4), adrenaline receptor β3 (ADRB3), AFP, anaplastic lymphoma kinase (ALK), androgen receptor, angiopoietin-binding cell surface receptor 2 (TIP). 2) Anti-desmosome 1 (Dsg1) autoantibody, anti-desmosome 3 (Dsg3) autoantibody, B7H3 (CD276), biotin, bone marrow stromal cell antigen 2 (BST2), BST1 / CD157, cancer / testis antigen 1 (NY-ESO-1), cancer / testis antigen 2 (LAGE-la), carbonic anhydrase IX (CAIX), carcinoembryonic antigen (CEA), CCCTC binding factor (zinc finger protein)-like (BORIS or imprinted site regulator sibling factor), CCR4, CD5, CD19, CD20, CD22, CD24, CD30, CD32 (FCG) R2A), CD33, CD34, CD38, CD44v6, CD72, CD79a, CD79b, CD97, CD99, CD123, CD171, CD179a, CD179b-IGLll, CD200R, CD276 / B7H3, CD300 molecular-like family member f (CD300LF), CDH1-CD324, CDH6, CDH17, CDH19, chromosome X open reading frame 61 (CXORF61), sealing protein 6 (CLDN6), sealing protein 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-α), 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-bike module 2 containing mucin-like hormone receptor-like (EMR2), Mutated elongation factor 2 (ELF2M), Ephrin B2, Ephrin type A receptor 2 (EphA2), Epidermal growth factor receptor (EGFR), Epidermal growth factor receptor variant III (EGFRviii), Epithelial cell adhesion molecule (EPCAM), ERG, ETS translocation-variant gene 6 located on chromosome 12p (ETV6-AML), Fc fragment of IgA receptor (FCAR or CD89), Fc receptor like 5 (FCRL5), Fibroblast activation 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-GMl, 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(l-4)bDGlcp(l-l)Cer), Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l-4)bDGlcp(l-l)Cer), GD3, GFRa4, Glycoprotein 100 (gp100), Glypican-3 (GPC3), Gonadotropin-releasing hormone receptor (CGHR or GR), GpA33, GpNMB, GPRC5D, Guanylate cyclase C (GCC), Mutated heat shock protein 70-2 (mut hsp70-2), Hepatitis A virus cellular receptor 1 (HAVCR1), Hexose portion of globoH glycosphingolipid (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, HTLVl-Tax, Human papillomavirus E6 (HPV E6), Human papillomavirus E7 (HPV E7), Human telomerase reverse transcriptase (hTERT), IgE, IL13Ra2, ILl lRa, Immunoglobulin lambda-like polypeptide 1 (IGLL1), Influenza A hemagglutinin (HA), Insulin-like growth factor 1 receptor (IGF-I receptor), Interleukin 11 receptor alpha (IL-llRa), Interleukin 13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2), Intestinal carboxylesterase, KIT (CD117), KSHVK8.1, KSHV-gH, LAMP1, Legumain, 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, Livl, Locus K 9 (LY6K), Low conductance chloride ion channel, Lymphocyte antigen 6 complex, Lymphocyte antigen 75 (LY75), Lymphocyte-specific protein tyrosine kinase (LCK), Mammary gland differentiation antigen (NY-BR-1), Melanoma antigen recognized by T cells 1 (Melan A or MARTI), Melanoma-associated antigen 1 (MAGE-A1), Melanoma cancer testis antigen-1 (MAD-CT-1), Melanoma cancer testis antigen-2 (MAD-CT-2), Melanoma cell apoptosis inhibitor (ML-IAP), Mesothelin, MPL, Mucin 1, cell surface associated (MUC1), N-Acetylglucosaminyltransferase V (NA17), Nectin-4, Neural cell adhesion molecule (NCAM), NKG2D, NYBR1, O-Acetyl-GD2 ganglioside (OAcGD2), Olfactory receptor 51E2 (OR51E2), Oncogenic fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl), P53 mutant, 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-beta), Polysialic acid, Pre-migratory trophoblast antigen sp32 (OY-TES1), Prostase, Prostate carcinoma tumor antigen-1 (PCTA-1 or Galectin 8), Prostate stem cell antigen (PSCA), Prostate-specific membrane antigen (PSMA), Prostate acid phosphatase (PAP), Prostein, Serine protease 21 (Testisin or PRSS21), Proteasome (Prosome Macropain) subunit beta9, PTK7, Ras G12V, RhoC, rat sarcoma (Ras) mutant, Receptor for advanced glycation end products (RAGE-1), Receptor tyrosine kinase like orphan receptor 1 (ROR1), Receptor tyrosine-protein kinase ERBB2 or Her-2 / neu, ubiquitous 1 (RU1), ubiquitous 2 (RU2), sarcoma translocation breakpoints, serine 2 (TMPRSS2) ETS fusion gene, sialyl Lewis adhesion molecule (sLe), SLAMF4, SLAMF6, Slea (CA19.9 or sialyl Lewis antigen), sperm protein 17 (SPA17), SART3, stage-specific embryonic antigens-4 (SSEA-4), STEAP1, survivin, SSX2, TCR gamma alternate 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, TNT antibody-recognized antigen, thyroid stimulating hormone receptor (TSHR), Timl- / HVCR1, tissue factor 1 (TF1), Tn 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 (TEM7R), tumor protein p53 (p53), tumor-associated glycoprotein 72 (TAG72), tyrosinase, tyrosinase-related protein 2 (TRP-2), UPK2, VEGFR2, V-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN), Wilms tumor protein (WT1), or X antigen family member 1A (XAGE1).

30. The viral particle of claim 29, wherein the target binding domain (T) binds to CD7.

31. The viral particle of claim 30, wherein the polypeptide comprises a heavy chain and a light chain comprising: a heavy chain variable region of the heavy chain having at least 90% sequence identity to SEQ ID NO: 36, and a light chain variable region of the light chain having at least 90% sequence identity to SEQ ID NO: 37, wherein the polypeptide comprises a sequence of HCDR1 as set forth in SEQ ID NO: 30; a sequence of HCDR2 as set forth in SEQ ID NO: 31; a sequence of HCDR3 as set forth in SEQ ID NO: 32; a sequence of LCDR1 as set forth in SEQ ID NO: 33; a sequence of LCDR2 as set forth in SEQ ID NO: 34; and a sequence of LCDR3 as set forth in SEQ ID NO:

35.

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

39.

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

34. The viral particle of 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 viral particle of claims 1- wherein the viral particle further comprises a nucleic acid molecule encoding a heterologous molecule of interest, wherein the heterologous molecule of interest is an siRNA, shRNA, non-coding RNA (e.g., a guide RNA for a CRISPR system), a peptide, a polypeptide, a protein, a viral payload, a viral genome, or a combination thereof.

36. A method of infecting a cell, the method comprising contacting the cell with a viral particle according to any one of claims 1 to 35.

37. A method of infecting a cell of a subject, the method comprising administering to the subject a pharmaceutical composition comprising a viral particle according to any one of claims 1 to 35.

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

39. The method of 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 of claim 38 or 39, wherein the edited target nucleic acid molecule is an endogenous gene.

41. The method of any one of claims 36 to 40, wherein the cell is a T cell, a B cell, a NK cell, a dendritic cell, a neutrophil, a macrophage, or a cancer cell.

42. A method of treating a disease or condition in a subject in need thereof, the method comprising administering to the subject a viral particle according to any one of claims 1 to 35, wherein the nucleic acid molecule encoding the gene editing system is expressed in the subject and treats the disease or condition in the subject.

43. A method of editing a target nucleic acid molecule in a cell, the method comprising contacting the cell with a viral particle comprising: (a) a VSV-G polypeptide comprising an amino acid sequence that is at least 70% identical to SEQ ID NO: 2 and comprising 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 a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the 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, a CD3+ T cell, a CD4+ T cell, a CD7+ T cell, or a CD8+ T cell; wherein the nucleic acid molecule encoding the gene editing system is expressed in the cell and edits the target nucleic acid molecule in the cell.

44. The method of claim 43, wherein the cell is a T cell, a CD3+ T cell, a CD4+ T cell, a CD7+ T cell, a CD8+ T cell, or a CD7+ NK cell.

45. 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 heterologous viral glycoprotein comprises a sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, and SEQ ID NO: 53; wherein the targeting moiety comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety; wherein the target binding domain comprises a sequence selected from the group consisting of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, and SEQ ID NO: 51; wherein the stalk moiety S1 comprises 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; wherein 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; wherein the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A; wherein 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; and wherein the variant Fc protein further comprises a transmembrane domain comprising a sequence selected from the group consisting of SEQ ID NO: 61 and SEQ ID NO: 62; and wherein the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule.

46. 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 heterologous viral glycoprotein comprises a sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, and SEQ ID NO: 53; wherein the targeting moiety comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety; wherein the target binding domain comprises a sequence selected from the group consisting of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, and SEQ ID NO: 51; wherein the stalk moiety S1 comprises the formula L1-Fc-L2-X1, wherein: L1 is a linker comprising a sequence selected from 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; 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; wherein 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; wherein the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A; wherein 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 a sequence selected from 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 X1is a polypeptide having the formula ECD-T M - a polypeptide of the ICD comprising a transmembrane domain, wherein: ECD is an extracellular domain having a sequence selected from SEQ ID NO: 59 or SEQ ID NO: 60 or a fragment thereof or is absent; T M is a transmembrane domain having the sequence of SEQ ID NO: 61 or SEQ ID NO: 62, or a fragment thereof; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, wherein the ICD comprises an env incorporation motif comprising an amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64 or ICD is absent; and wherein the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule.

47. 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 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; wherein the targeting moiety comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety; wherein the target binding domain comprises a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51 ; wherein the stalk moiety 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; wherein 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; wherein the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A; wherein 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 X1is a polypeptide having the formula ECD-T M - a polypeptide of the ICD comprising a transmembrane domain, wherein: ECD is an extracellular domain having the sequence of SEQ ID NO: 60 or a fragment thereof or is absent; T M is a transmembrane domain having the sequence of SEQ ID NO: 62 or a fragment thereof; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, wherein the ICD comprises an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64 or ICD is absent; and wherein the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule.

48. 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 heterologous viral glycoprotein comprises the sequence of SEQ ID NO: 23 or SEQ ID NO: 25; wherein the targeting moiety comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety; wherein the target binding domain comprises the sequence of SEQ ID NO: 39; wherein the stalk moiety S1 comprises the formula L1-Fc-L2-X1, wherein: 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; X1is a polypeptide having the formula ECD-T M - a polypeptide of the ICD comprising a transmembrane domain, wherein: ECD is an extracellular domain having the sequence of SEQ ID NO: 60; T M is a transmembrane domain having the sequence of SEQ ID NO: 62; ICD is a cell internalization domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, wherein the ICD comprises an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63; and wherein the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule.

49. 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 heterologous viral glycoprotein comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identical to SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identical to SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identical to SEQ ID NO: 23 or SEQ ID NO: 25; the targeting moiety comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 83, at least 95% identical to SEQ ID NO: 83, at least 99% identical to SEQ ID NO: 83, or at least 100% identical to SEQ ID NO: 83; and the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule. the heterologous viral glycoprotein comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identical to SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identical to SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identical to SEQ ID NO: 23 or SEQ ID NO: 25; the targeting moiety comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 83, at least 95% identical to SEQ ID NO: 83, at least 99% identical to SEQ ID NO: 83, or at least 100% identical to SEQ ID NO: 83; and the nucleic acid molecule encoding a gene editing system encodes a CRISPR-Cas system comprising a Cas9 protein and a guide RNA, a single guide RNA, or at least one nucleic acid molecule that targets the Cas protein to a target nucleic acid molecule.

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