Nucleic acid constructs and engineered cells for allogeneic therapies

Abstract

The present disclosure relates to nucleic acid molecules that include both CAR-encoding sequences and one or more shRNA sequences in a single nucleic acid molecule, as well as cells and cell-based therapies that incorporate and express such nucleic acid molecules. Certain aspects of the disclosure therefore also relate to engineered cells for use in cell therapies, e.g., allogeneic cell therapies.

Description

[0001]NUCLEIC ACID CONSTRUCTS AND ENGINEERED CELLS FOR ALLOGENEIC THERAPIES CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S. Provisional Application No 63 / 510,825, filed June 28, 2023. The entire contents of the above-identified application are hereby fully incorporated herein by reference. SEQUENCE LISTING This application contains a sequence listing in electronic form as an XML file entitled 1004034_0011_SequenceListing with size 136,842 bytes created on June 28, 2024. The content of the sequence listing is incorporated herein in its entirety. FIELD OF THE INVENTION The invention relates generally to synthetic nucleic acid molecules and cell-based therapies. BACKGROUND OF THE INVENTION With recent FDA approval of the first chimeric antigen receptor (CAR) T-cell therapy (Wall Street Journal, August 30, 2017), certain cell-based immunotherapeutics have begun to realize their clinical potential. However, therapies based on autologous T cells confront severe limitations, mainly related to the fact that the product has to be generated from each patient's cells, in a time-consuming and costly process, and with the risk of manufacturing failure (Zhao et al. J Hematol Oncol. (2018) 11:132). A need exists for nucleic acid constructs and genetically altered cells capable of providing CAR-expressing cell populations for use in allogeneic therapies. The present disclosure addresses this need. BRIEF SUMMARY OF THE INVENTION The current disclosure relates, at least in part, to discovery and development of nucleic acid molecules capable of driving genetic alterations of mammalian cells to create engineered, clinically useful CAR-expressing cells for use in allogeneic therapies. In certain aspects, the nucleic acid molecules and CAR-expressing cells of the disclosure are rendered safe and persistent for activity in a recipient, non-self mammal by encoding for shRNAs capable of downregulating, e.g., T cell receptor (TCR) complexes, MHC class I molecules, and MHC class II molecules, optionally among others, in nucleic acid construct-containing cells. In some aspects, terminal repeat-recognizing mobile element enzyme(s) and / or transposase(s) effect genomic integration of the nucleic acid molecules of the disclosure. In one aspect, the disclosure provides a nucleic acid molecule that includes: (a) a type III RNA polymerase III promoter operably linked to a sequence encoding for a short hairpin RNA (shRNA), where the shRNA is capable of annealing and reducing expression of a messenger RNA (mRNA) that is a MHC class I gene mRNA, a MHC class II gene mRNA and / or a TCRα chain mRNA; and (b) two terminal repeat sequences capable of recognition by an enzyme capable of performing targeted genomic integration of the nucleic acid molecule, where the enzyme is derived from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Myotis lucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, troglodytes, Molossus molossus, or Homo sapiens. In one embodiment, the nucleic acid molecule further includes a sequence encoding for a chimeric antigen receptor (CAR). Optionally, the chimeric antigen receptor is an anti-CD19 CAR. Optionally, the anti-CD19 CAR is operably linked to a eukaryotic translation elongation factor 1 α (EF-1α) promoter. In another embodiment, the CAR includes an anti-CD19 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence. In certain embodiments, the nucleic acid molecule includes two terminal repeat sequences. In one embodiment, the nucleic acid molecule includes one of the following ordered series of sequences: (a) terminal repeat sequence – EF1α promoter operably linked to a CAR-encoding sequence – U6 promoter operably linked to a shRNA – terminal repeat sequence; (b) terminal repeat sequence – U6 promoter operably linked to a shRNA – EF1α promoter operably linked to a CAR- encoding sequence – terminal repeat sequence, where the shRNA and the CAR-encoding sequence are transcribed in the same direction; (c) terminal repeat sequence – U6 promoter operably linked to a shRNA – EF1α promoter operably linked to a CAR-encoding sequence – terminal repeat sequence, where the shRNA and the CAR-encoding sequence are transcribed in opposite directions; or (d) terminal repeat sequence – EF1α promoter operably linked to a CAR-encoding sequence – U6 promoter operably linked to two or more shRNA sequences – terminal repeat sequence. In another embodiment, the CAR-encoding sequence encodes for an anti-CD19 CAR. Optionally, the CAR includes an anti-CD19 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence. In some embodiments, the CAR-encoding sequence encodes for an anti-CD20 CAR. Optionally, the CAR includes an anti-CD20 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence. In certain embodiments, the CAR-encoding sequence encodes for an anti-CD22 CAR. Optionally, the CAR includes an anti-CD22 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence. In another embodiment, the CAR-encoding sequence encodes for an anti-B7 H3 CAR. Optionally, the CAR includes an anti-B7 H3 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB or CD28 signaling domain sequence and a CD3 zeta signaling domain sequence. In other embodiments, the CAR-encoding sequence encodes for an anti-human epidermal growth factor receptor 2 (Her2) CAR. Optionally, the CAR includes an anti-Her2 FMC light chain, a CD8 hinge and transmembrane domain, a CD28 signaling domain sequence and a CD3 zeta signaling domain sequence. In one embodiment, the shRNA is a Beta-2-microglobulin-targeting shRNA. In another embodiment, the shRNA is a T cell receptor alpha constant (TRAC)-targeting shRNA. In an additional embodiment, the shRNA is a Class II Major Histocompatibility Complex Transactivator (CIITA)-targeting shRNA. In one embodiment, the shRNA is a transporter 1, ATP binding cassette subfamily B member (TAP1)-targeting shRNA. In another embodiment, the shRNA is a transporter 2, ATP binding cassette subfamily B member (TAP2)-targeting shRNA. In certain embodiments, the shRNA is a CD3ζ-targeting shRNA. In some embodiments, the nucleic acid molecule includes one or more of SEQ ID NOs: 1-5, 12-15, 57, 58 and / or 76-80. In certain embodiments, the nucleic acid molecule includes a linker sequence between sequences encoding for shRNAs. Optionally, the linker sequence includes one or more of SEQ ID NOs: 59-65. In some embodiments, the enzyme capable of performing targeted genomic integration is Tn1, Tn2, Tn3, Tn5, Tn7, Tn9, Tn10, Tn552, Tn903, Tn1000 / Gamma-delta, Tn / O, tnsA, tnsB, tnsC, tniQ, IS10, ISS, IS911, Minos, Sleeping beauty, piggyBac, Tol2, Mos1, Himar1, Hermes, Tol2, Minos, Tel, P-element, MuA, Ty1, Chapaev, transib, Tc1 / mariner, or Tc3 donor DNA system. In another embodiment, the enzyme capable of performing targeted genomic integration includes an amino acid sequence of SEQ ID NOs: 66, 69-75, and variants thereof. In one embodiment, the two terminal repeat sequences include: a Myotis lucifugus mobile element enzyme (MLT) left terminal end (SEQ ID NO: 67), or a sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto; and / or a MLT right terminal end (SEQ ID NO: 68), or a sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto. Optionally, the nucleic acid molecule includes both the MLT left terminal end and the MLT right terminal end sequences. In another embodiment, the nucleic acid molecule further includes a rabbit β-globin polyadenylation signal (rBG PA). An additional aspect of the disclosure provides a cell harboring the nucleic acid molecule of the instant disclosure. In certain embodiments, the cell further includes the enzyme capable of performing targeted genomic integration of the nucleic acid molecule. Optionally, the enzyme capable of performing targeted genomic integration of the nucleic acid molecule is encoded by an RNA. In some embodiments, the cell is an immune cell. Optionally, the cell is a T cell, a macrophage, or a natural killer (NK) cell. In certain embodiments, the cell further includes an exogenous copy of CD47 and / or HLA- E. Optionally, the exogenous copy of CD47 and / or HLA-E is integrated in the genome of the cell. In one embodiment, the cell is administered to a subject. Optionally, the cell is administered to a mammalian subject. Optionally, the cell is administered to a human. In certain embodiments, the cell is a component of an allogeneic therapy. Another aspect of the disclosure provides a method of treating a hematological cancer in a subject, the method involving administering a cell of the disclosure to the subject, thereby treating the hematological cancer in the subject. In certain embodiments, the hematological cancer is leukemia, lymphoma, or multiple myeloma. A further aspect of the disclosure provides a method of making a chimeric antigen receptor (CAR)-immune cell, the method involving: obtaining an immune cell from a sample obtained from a subject, the sample including immune cells or immune cell progenitors; and transfecting the immune cell with a first nucleic acid molecule of the disclosure and a second nucleic acid molecule encoding the enzyme capable of performing targeted genomic integration of the first nucleic acid molecule (optionally, the second nucleic acid molecule is RNA), to thereby create a transfected CAR-immune cell, where the immune cell is a T cell, a macrophage, or a natural killer (NK) cell. Another aspect of the disclosure provides a method of making a chimeric antigen receptor (CAR)-immune cell, the method involving: administering to a subject a first nucleic acid molecule of the instant disclosure and a second nucleic acid molecule encoding the enzyme capable of performing targeted genomic integration of the first nucleic acid molecule (optionally, the second nucleic acid molecule is RNA), where said administering uses lipid nanoparticles (LNPs) capable of directing the first and second nucleic acid molecules to an immune cell, to thereby create a CAR- immune cell, where the immune cell is a T cell, a macrophage, or a natural killer (NK) cell. In one embodiment, the immune cell is a T cell. In another embodiment, the transfected CAR-immune cell is a CAR-T cell. Optionally, the CAR-T cell is an autologous CAR-T cell or an allogeneic CAR-T cell. In some embodiments, the immune cell is a macrophage. In a related embodiment, the transfected CAR-immune cell is a CAR-M cell. Optionally, the transfected CAR-immune cell is an autologous CAR-M cell or an allogeneic CAR-M cell. In certain embodiments, the immune cell is a natural killer (NK) cell. In a related embodiment, the transfected CAR-immune cell is a CAR-NK cell. Optionally, the transfected CAR-immune cell is an autologous CAR-NK cell or an allogeneic CAR-NK cell. A further aspect of the disclosure provides a cell having a genomic integrant of a nucleic acid of the disclosure, where the genomic integrant is prepared using a Myotis lucifugus mobile element enzyme (MLT), to drive the genomic integration event. In a related embodiment, the genomic integrant is located at least 1 kb away from any promoter start site within the genome of the cell. Definitions Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. “About” can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. In certain embodiments, the term "approximately" or "about" refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). Unless otherwise clear from context, all numerical values provided herein are modified by the term “about.” The term “administration” refers to introducing a substance into a subject. In general, any route of administration may be utilized including, for example, parenteral (e.g., intravenous), oral, topical, subcutaneous, peritoneal, intra-arterial, inhalation, vaginal, rectal, nasal, introduction into the cerebrospinal fluid, or instillation into body compartments. In some embodiments, administration is parenteral. In some embodiments, administration is intravenous. The term “antigen” or “Ag” as used herein is defined as a molecule that can be targeted by an antibody or antibody fragment thereof. As used herein, a “tumor antigen” means a biological molecule having antigenicity, expression of which is associated with a neoplastic cell. The tumor antigens targeted in the present disclosure include a tumor specific antigen (an antigen which is present only in tumor cells and is not found in other normal cells), and a tumor-associated antigen (an antigen which is also present in other organs and tissues or heterogeneous and allogeneic normal cells, or an antigen which is expressed on the way of development and differentiation). As used herein, the term “autologous” is meant to refer to any material derived from the same individual to which it is later to be re-introduced into the individual. As used herein “endogenous” refers to any material from or produced inside an organism, cell, tissue or system. A “co-stimulatory molecule” refers to the cognate binding partner on a T-cell that specifically binds with a co-stimulatory ligand, thereby mediating a co-stimulatory response by the T-cell, such as, but not limited to, proliferation. Co-stimulatory molecules include, but are not limited to an MHC class I molecule, BTLA and a Toll ligand receptor. A “co-stimulatory signal”, as used herein, refers to a signal, which in combination with a primary signal, such as TCR / CD3 ligation, leads to T-cell proliferation, activation, and / or upregulation or downregulation of key molecules. By “control” or “reference” is meant a standard of comparison. In one aspect, as used herein, “changed as compared to a control” sample or subject is understood as having a level that is statistically different than a sample from a normal, untreated, or control sample. Control samples include, for example, cells in culture, one or more laboratory test animals, or one or more human subjects. Methods to select and test control samples are within the ability of those in the art. Determination of statistical significance is within the ability of those skilled in the art, e.g., the number of standard deviations from the mean that constitute a positive result. The term “cancer” refers to a malignant neoplasm (Stedman's Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990). Exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary cancer (e.g., cholangiocarcinoma); bladder cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma); connective tissue cancer; epithelial carcinoma; ependymoma; endotheliosarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma); endometrial cancer (e.g., uterine cancer, uterine sarcoma); esophageal cancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarcinoma); Ewing's sarcoma; ocular cancer (e.g., intraocular melanoma, retinoblastoma); familiar hypereosinophilia; gall bladder cancer; gastric cancer (e.g., stomach adenocarcinoma); gastrointestinal stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)); hematopoietic cancers (e.g., leukemia such as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL)); lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., Waldenström's macroglobulinemia), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T- cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, and anaplastic large cell lymphoma); a mixture of one or more leukemia / lymphoma as described above; and multiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease); hemangioblastoma; hypopharynx cancer; inflammatory myofibroblastic tumors; immunocytic amyloidosis; kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma); liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorder (MPD) (e.g., polycythemia vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)); neuroblastoma; neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis); neuroendocrine cancer (e.g., gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor); osteosarcoma (e.g., bone cancer); ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma); papillary adenocarcinoma; pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), Islet cell tumors); penile cancer (e.g., Paget's disease of the penis and scrotum); pinealoma; primitive neuroectodermal tumor (PNT); plasma cell neoplasia; paraneoplastic syndromes; intraepithelial neoplasms; prostate cancer (e.g., prostate adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer; skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small bowel cancer (e.g., appendix cancer); soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland carcinoma; small intestine cancer; sweat gland carcinoma; synovioma; testicular cancer (e.g., seminoma, testicular embryonal carcinoma); thyroid cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer); urethral cancer; vaginal cancer; and vulvar cancer (e.g., Paget's disease of the vulva). A "chimeric antigen receptor" or "CAR" as used herein refers to a chimeric receptor (i.e. composed of parts from different sources) that has at least a binding moiety with a specificity for an antigen (which can e.g. be derived from an antibody, a receptor or its cognate ligand) and a signaling moiety that can transmit a signal in an immune cell (e.g. a CD3 zeta chain. Other signaling or cosignaling moieties can also be used, such as e.g. a Fc epsilon Rl gamma domain, a CD3 epsilon domain, the recently described DAP10 / DAP12 signaling domain, or domains from CD28, 4-1BB, 0X40, ICOS, DAP10, DAP12, CD27, and CD2 as costimulatory domain). A "chimeric NK receptor" is a CAR wherein the binding moiety is derived or isolated from a NK receptor. A "TCR" as used herein refers to a T cell receptor. In the context of adoptive cell transfer, this typically refers to an engineered TCR, i.e. a TCR that has been engineered to recognize a specific antigen, most typically a tumor antigen. An "endogenous TCR" as used herein refers to a TCR that is present endogenously, on non-modified cells (typically T cells). The TCR is a disulfide-linked membrane-anchored heterodimeric protein normally consisting of the highly variable alpha (a) and beta (b) chains expressed as part of a complex with the invariant CD3 chain molecules. The TCR receptor complex is an octomeric complex of variable TCR receptor a and b chains with the CD3 co-receptor (containing a CD3y chain, a CD36 chain, and two CD3e chains) and two CD3 z chains (aka CD247 molecules). The term "functional TCR" as used herein means a TCR capable of transducing a signal upon binding of its cognate ligand. Typically, for allogeneic therapies, engineering will take place to reduce or impair the TCR function, e.g. by knocking out or knocking down at least one of the TCR chains. An endogenous TCR in an engineered cell is considered functional when it retains at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, or even at least 90% of signalling capacity (or T cell activation) compared to a cell with endogenous TCR without any engineering. Assays for assessing signalling capacity or T cell activation are known to the person skilled in the art, and include amongst others an ELISA measuring interferon gamma. According to alternative embodiments, an endogenous TCR is considered functional if no engineering has taken place to interfere with TCR function. The term "immune cells" as used herein refers to cells that are part of the immune system (which can be either the adaptive or the innate immune system). Immune cells as used herein are typically immune cells that are manufactured for adoptive cell transfer (either autologous transfer or allogeneic transfer). Many different types of immune cells are used for adoptive therapy and thus are envisaged for use in the methods described herein. Examples of immune cells include, but are not limited to, T cells, NK cells, NKT cells, lymphocytes, dendritic cells, myeloid cells, stem cells, progenitor cells or iPSCs. The latter three are not immune cells as such, but can be used in adoptive cell transfer for immunotherapy (see e.g. Jiang et al., Cell Mol Immunol 2014; Themeli et al., Cell Stem Cell 2015). Typically, while the manufacturing starts with stem cells or iPSCs (or may even start with a dedifferentiation step from immune cells towards iPSCs), manufacturing will entail a step of differentiation to immune cells prior to administration. Stem cells, progenitor cells and iPSCs used in manufacturing of immune cells for adoptive transfer (i.e., stem cells, progenitor cells and iPSCs or their differentiated progeny that are transduced with a CAR as described herein) are considered as immune cells herein. According to particular embodiments, the stem cells envisaged in the methods do not involve a step of destruction of a human embryo. Particularly envisaged immune cells include white blood cells (leukocytes), including lymphocytes, monocytes, macrophages and dendritic cells. Particularly envisaged lymphocytes include T cells, NK cells and B cells, most particularly envisaged are T cells. In the context of adoptive transfer, immune cells can be primary cells (i.e. cells isolated directly from human or animal tissue, and not or only briefly cultured), or in certain embodiments can be passaged / expanded cells, particularly for allogeneic therapies described herein. The terms "domain" and "motif, used interchangeably herein, refer to both structured domains having one or more particular functions and unstructured segments of a polypeptide that, although unstructured, retain one or more particular functions. For example, a structured domain may encompass but is not limited to a continuous or discontinuous plurality of amino acids, or portions thereof, in a folded polypeptide that comprise a three-dimensional structure which contributes to a particular function of the polypeptide. In other instances, a domain may include an unstructured segment of a polypeptide comprising a plurality of two or more amino acids, or portions thereof, that maintains a particular function of the polypeptide unfolded or disordered. Also encompassed within this definition are domains that may be disordered or unstructured but become structured or ordered upon association with a target or binding partner. Non-limiting examples of intrinsically unstructured domains and domains of intrinsically unstructured proteins are described, e.g., in Dyson & Wright. Nature Reviews Molecular Cell Biology 6: 191-208. By "fragment" is meant a portion, e.g., a portion of a polypeptide or nucleic acid molecule. This portion contains, preferably, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the entire length of the reference nucleic acid molecule or polypeptide. For example, a fragment may contain 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 nucleotides or amino acids. However, the disclosure also comprises polypeptides and nucleic acid fragments, so long as they exhibit the desired / indicated biological activity / activities of the full length polypeptides and nucleic acid, respectively. A nucleic acid fragment of almost any length is employed. For example, illustrative polynucleotide segments with total lengths of about 10,000, about 5000, about 3000, about 2,000, about 1,000, about 500, about 200, about 100, about 50 base pairs in length (including all intermediate lengths) are included in many implementations of this disclosure. Similarly, a polypeptide fragment of almost any length is employed. For example, illustrative polypeptide segments with total lengths of about 10,000, about 5,000, about 3,000, about 2,000, about 1,000, about 5,000, about 1,000, about 500, about 200, about 100, or about 50 amino acids in length (including all intermediate lengths) are included in many implementations of this disclosure. “Heterologous," as used herein, means a nucleotide or polypeptide sequence that is not found in the native (e.g., naturally-occurring) nucleic acid or protein, respectively. As used herein, the term "immune cells" generally includes white blood cells (leukocytes) which are derived from hematopoietic stem cells (HSC) produced in the bone marrow. "Immune cells" includes, e.g., lymphocytes (T cells, B cells, natural killer (NK) cells) and myeloid-derived cells (neutrophil, eosinophil, basophil, monocyte, macrophage, dendritic cells). "T cell" includes all types of immune cells expressing CD3 including T-helper cells (CD4+cells), cytotoxic T-cells (CD8+cells), T-regulatory cells (Treg) and gamma-delta T cells. A "cytotoxic cell" includes CD8+T cells, natural-killer (NK) cells, and neutrophils, which cells are capable of mediating cytotoxicity responses. As used herein, the term "stem cell" generally includes pluripotent or multipotent stem cells. "Stem cells" includes, e.g., embryonic stem cells (ES); mesenchymal stem cells (MSC); induced- pluripotent stem cells (iPS); and committed progenitor cells (hematopoietic stem cells (HSC); bone marrow derived cells, neural progenitor cells, etc.). As used herein, the term "heteromeric" refers to a polypeptide or protein that contains more than one kind of subunit. Such heteromeric polypeptides may, in some instances, be referred to as "a heteromer". Heteromeric polypeptides may contain two or more different polypeptides, wherein different polypeptides are defined at least as two polypeptides that are not identical, however, such different polypeptides may or may not include one or more portions of similar and / or identical amino acid sequence. In some instances, the two or more polypeptides of a heteromer share no identical amino acid sequence or share no identical domains. A heteromer may, in some instances, consist of two different polypeptides or two different types of polypeptides and may be referred to as a heterodimer. In some instances, a heteromer may consist of three different polypeptides or three different types of polypeptides and may be referred to as a heterotrimer. In some instances, a heteromer may consist of two or more different polypeptides or two or more different types of polypeptides, including but not limited to, e.g., three or more different polypeptides, four or more different polypeptides, five or more different polypeptides, six or more different polypeptides, seven or more different polypeptides, eight or more different polypeptides, etc. The phrase "graft versus host disease" or "GvHD" refers to a condition that might occur after an allogeneic transplant. In GvHD, the donated bone marrow, peripheral blood (stem) cells or other immune cells view the recipient's body as foreign, and the donated cells attack the body. As donor immunocompetent immune cells, such as T cells, are the main driver for GvHD, one strategy to prevent GvHD is by reducing (TCR-based) signaling in these immunocompetent cells, e.g. by directly or indirectly inhibiting the function of the TCR complex. The phrase "adoptive cellular therapy", "adoptive cell transfer", or "ACT" as used herein refers to the transfer of cells, most typically immune cells, into a subject (e.g., a patient). These cells may have originated from the subject (in case of autologous therapy) or from another individual (in case of allogeneic therapy). The goal of the therapy is to improve immune functionality and characteristics, and in cancer immunotherapy, to raise an immune response against the cancer. Although T cells are most often used for ACT, it is also applied using other immune cell types such as NK cells, lymphocytes (e.g. tumor-infiltrating lymphocytes (TILs)), dendritic cells and myeloid cells. The terms “isolated,” “purified,” or “biologically pure” refer to material that is free to varying degrees from components which normally accompany it as found in its native state. “Isolate” denotes a degree of separation from original source or surroundings. “Purify” denotes a degree of separation that is higher than isolation. As used herein, "neoplasia" means a disease state of a human or an animal in which there are cells and / or tissues which proliferate abnormally. Neoplastic conditions include, but are not limited to, cancers, sarcomas, tumors, leukemias, lymphomas, and the like. A neoplastic condition refers to the disease state associated with the neoplasia. Hepatocellular carcinoma, colon cancer (e.g., colorectal cancer), lung cancer and ovarian cancer are examples (non-limiting) of a neoplastic condition. A "cancer" in a subject refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Often, cancer cells will be in the form of a tumor, but such cells may exist alone within a subject, or may be a non- tumorigenic cancer cell, such as a leukemia cell. Examples of cancer include but are not limited to hepatic carcinoma, colon cancer, colorectal cancer, breast cancer, a melanoma, adrenal gland cancer, biliary tract cancer, bladder cancer, brain or central nervous system cancer, bronchus cancer, blastoma, carcinoma, a chondrosarcoma, cancer of the oral cavity or pharynx, cervical cancer, esophageal cancer, gastrointestinal cancer, glioblastoma, hepatoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreas cancer, peripheral nervous system cancer, prostate cancer, sarcoma, salivary gland cancer, small bowel or appendix cancer, small-cell lung cancer, squamous cell cancer, stomach cancer, testis cancer, thyroid cancer, urinary bladder cancer, uterine or endometrial cancer, and vulval cancer. "Operably linked" refers to a juxtaposition wherein the components so described are in a relationship permitting them to function in their intended manner. For instance, a promoter is operably linked to a coding sequence if the promoter affects its transcription or expression. The term "nucleic acid molecule" synonymously referred to as "nucleotides" or "nucleic acids" or "polynucleotide" as used herein refers to any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. Nucleic acid molecules include, without limitation, single- and double-stranded DNA, DNA that is a mixture of single- and double- stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single- stranded or, more typically, double-stranded or a mixture of single- and double- stranded regions. In addition, "polynucleotide" refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA. The term polynucleotide also includes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications may be made to DNA and RNA; thus, "polynucleotide" embraces chemically, enzymatically or metabolically modified forms of polynucleotides as typically found in nature, as well as the chemical forms of DNA and RNA characteristic of viruses and cells. "Polynucleotide" also embraces relatively short nucleic acid chains, often referred to as oligonucleotides. The term "exogenous" as used herein, particularly in the context of cells or immune cells, refers to any material that is present and active in an individual living cell but that originated outside that cell (as opposed to an endogenous factor). The phrase "exogenous nucleic acid molecule" thus refers to a nucleic acid molecule that has been introduced in the (immune) cell, typically through transduction or transfection. The term "endogenous" as used herein refers to any factor or material that is present and active in an individual living cell and that originated from inside that cell (and that are thus typically also manufactured in a non-transduced or non-transfected cell). "Isolated" as used herein means a biological component (such as a nucleic acid, peptide or protein) has been substantially separated, produced apart from, or purified away from other biological components of the organism in which the component naturally occurs, i.e., other chromosomal and extrachromosomal DNA and RNA, and proteins. Nucleic acids, peptides and proteins that have been "isolated" thus include nucleic acids and proteins purified by standard purification methods. "Isolated" nucleic acids, peptides and proteins can be part of a composition and still be isolated if such composition is not part of the native environment of the nucleic acid, peptide, or protein. The term also embraces nucleic acids, peptides and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids. "Multiplexed" as used herein in the context of gene editing refers to the simultaneous targeting of two or more (i.e. multiple) related or unrelated targets. The term "RNA interference molecule" as used herein refers to an RNA (or RNA-like) molecule that inhibits gene expression or translation, by neutralizing targeted mRNA molecules. Examples include siRNA (including shRNA) or miRNA molecules. "Multiplexed RNA interference molecules" as used herein thus are two or more molecules that are simultaneously present for the concomitant downregulation of one or more targets. Typically, each of the multiplexed molecules will be directed against a specific target, but two molecules can be directed against the same target (and can even be identical). A "promoter" as used herein is a regulatory region of nucleic acid usually located adjacent to a gene region, providing a control point for regulated gene transcription. A "multiplex" is a polynucleotide that encodes for a plurality of molecules of the same type, e.g., a plurality of siRNA or shRNA or miRNA. Within a multiplex, when molecules are of the same type (e.g., all shRNAs), they may be identical or comprise different sequences. Between molecules that are of the same type, there may be intervening sequences such as the linkers described herein. A multiplex may be single stranded, double stranded or have both regions that are single stranded and regions that are double stranded. The terms "polypeptide," "peptide," and "protein", used interchangeably herein, refer to a polymeric form of amino acids of any length, which can include genetically coded and non- genetically coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones. The term includes fusion proteins, including, but not limited to, fusion proteins with a heterologous amino acid sequence, fusions with heterologous and homologous leader sequences, with or without N-terminal methionine residues; immunologically tagged proteins; and the like. A "short hairpin RNA" or "small hairpin RNA" ("shRNA") is an artificial RNA molecule with a tight hairpin turn that can be used to silence target gene expression via RNA interference (RNAi). The successful expression of shRNA has largely been dependent upon coupling with a polymerase III (Pol III) promoter (e.g. HI, U6) that generate RNA species lacking a 5' cap and 3' polyadenylation, enabling processing of the shRNA duplex. Once transcribed, the shRNA undergoes processing, export from the nucleus, further processing and loading into the RNA-induced silencing complex (RISC) complex leading to the targeting degradation of mRNA of choice (Moore et al., 2010). Cells containing at least two RNA interference molecules can have advantages, particularly therapeutic benefits. RNA interference molecules can indeed be directed against targets of which (over)expression is undesirable. An "engineered cell" as used herein is a cell that has been modified through human intervention (as opposed to naturally occurring mutations). As used herein, the term "subject" includes humans and mammals (e.g., mice, rats, pigs, cats, dogs, and horses). In many embodiments, subjects are mammals, particularly primates, especially humans. In some embodiments, subjects are livestock such as cattle, sheep, goats, cows, swine, and the like; poultry such as chickens, ducks, geese, turkeys, and the like; and domesticated animals particularly pets such as dogs and cats. In some embodiments (e.g., particularly in research contexts) subject mammals will be, for example, rodents (e.g., mice, rats, hamsters), rabbits, primates, or swine such as inbred pigs and the like. As used herein, the term "tumor" means a mass of transformed cells that are characterized by neoplastic uncontrolled cell multiplication and at least in part, by containing angiogenic vasculature. The abnormal neoplastic cell growth is rapid and continues even after the stimuli that initiated the new growth has ceased. The term "tumor" is used broadly to include the tumor parenchymal cells as well as the supporting stroma, including the angiogenic blood vessels that infiltrate the tumor parenchymal cell mass. Although a tumor generally is a malignant tumor, i.e., a cancer having the ability to metastasize (i.e., a metastatic tumor), a tumor also can be nonmalignant (i.e., non-metastatic tumor). Tumors are hallmarks of cancer, a neoplastic disease the natural course of which is fatal. Cancer cells exhibit the properties of invasion and metastasis and are highly anaplastic. As used herein, the terms "treatment," "treating," "treat" and the like, refer to obtaining a desired pharmacologic and / or physiologic effect. The effect can be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or can be therapeutic in terms of a partial or complete cure for a disease and / or adverse effect attributable to the disease. "Treatment," as used herein, covers any treatment of a disease or condition in a mammal, particularly in a human, and includes: (a) preventing the disease from occurring in a subject which can be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease. An "effective amount" or “therapeutically effective amount” of an agent or cell therapy described herein is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to delay or minimize one or more symptoms associated with the condition. A therapeutically effective amount of an agent or cell therapy means an amount of therapeutic agent or cell, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms, signs, or causes of the condition, and / or enhances the therapeutic efficacy of another therapeutic agent. The transitional term “comprising,” which is synonymous with “including,” is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. By contrast, the transitional phrase “consisting of” excludes any element, step, or ingredient not specified in the claim. The transitional phrase “consisting essentially of” limits the scope of a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. A "vector" or "expression vector" is a replicon, such as plasmid, phage, virus, or cosmid, to which another DNA segment, i.e. an "insert", may be attached so as to bring about the replication of the attached segment in a cell. Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms "a", "an", and "the" are understood to be singular or plural. The phrase “pharmaceutically acceptable carrier” is art recognized and includes a pharmaceutically acceptable material, composition or vehicle, suitable for administering compounds of the present disclosure to mammals. The carriers include liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject agent from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen- free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another aspect includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it is understood that the particular value forms another aspect. It is further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. It is also understood that throughout the application, data are provided in a number of different formats and that this data represent endpoints and starting points and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed. Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 as well as all intervening decimal values between the aforementioned integers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to sub-ranges, “nested sub- ranges” that extend from either end point of the range are specifically contemplated. For example, a nested sub-range of an exemplary range of 1 to 50 may comprise 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in the other direction. Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All published foreign patents and patent applications cited herein are incorporated herein by reference. Genbank and NCBI submissions indicated by accession number cited herein are incorporated herein by reference. All other published references, documents, manuscripts and scientific literature cited herein are incorporated herein by reference. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. BRIEF DESCRIPTION OF THE DRAWINGS The following detailed description, given by way of example, but not intended to limit the disclosure solely to the specific embodiments described, may best be understood in conjunction with the accompanying drawings, in which: FIG. 1 shows a schematic diagram of exemplary modifications made to donor cells to generate a CD19+CAR-T cell for use, e.g., as an allogeneic cell therapy. Through the use of short hairpin RNAs (shRNAs), components of major histocompatibility complex 1 and / or 2 (MHC I and MHC II, respectively) as well as T-cell receptor alpha (TCRα) can be knocked down to reduce MHC and / or TCR activity. Additionally, cell surface receptor(s), such as CD47 (pictured), can also be knocked in, to protect cells from destruction by circulating macrophages. FIG.2 demonstrates an exemplary interaction of T-cell receptor (TCR) with HLA or MHC class I. A CD3ζ-targeting shRNA can be used to achieve knockdown of T-cell receptor alpha (TCRα). Knockdown / ablation of TCRα is a preferred method for decreasing TCR activity as a whole, because TCRα is encoded by a single gene, whereas TCRβ is encoded by two genes. FIG.3 shows a schematic diagram of the formation of major histocompatibility complex I (MCH-I) and its presentation on the cell surface. MHC-I is formed from cytosolic proteins which, through ubiquitination and proteasomal degradation, are processed into short component peptides. Such peptides are then transported by TAP 1 and 2 transporter proteins, through the endoplasmic reticulum (ER), and are shuttled via the Golgi apparatus to the cell surface, where presentation and interaction with CD8+T cells occurs. Here, acquired mutations in the antigen presentation pathway drive loss of MHC-I. FIG. 4 demonstrates the superior viability of HH cells (a mature T cell line derived from peripheral blood of a patient with aggressive cutaneous T cell leukemia / lymphoma) relative to THP1 cells (a spontaneously immortalized monocyte-like cell line, derived from the peripheral blood of a childhood case of acute monocytic leukemia), as a target cell line for shRNAs, for use in screening. At left, flow cytometry identified high cell surface levels in HH cells of both MHC class I and MHC class II proteins. The middle graph shows that levels of beta 2 microglobulin (B2M) mRNA in HH cells were nearly as high as GAPDH control mRNA levels, as measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). At right, HH cells were shown to transfect well (as demonstrated by intracellular accumulation of a transfected GFP marker) and also showed robust cellular viability post-transfection (delivery and viability levels exceeding 80% of cells in the transfected population were respectively observed). HH cells were therefore assessed as showing good transfection and cell growth. FIG.5 depicts a schematic diagram of various formats of shRNA constructs initially made and assessed for activity. FIG.6 shows a schematic diagram of the U6 promoter-shRNA junction of initial constructs (top strand: SEQ ID NO: 76; bottom strand: SEQ ID NO: 77), with sites of revisions numbered, accompanied by a table of such revisions and associated construct names tested in FIGs.7A and 7B below. FIGs.7A and 7B show construct shRNA-mediated MHC class I knockdown levels at day 7 in construct-transfected HH cells, using revised constructs ("NT" is a negative control; "OL021" and "OL022" are siRNA controls; "P217" and "P218" are shRNAs with reversed target / seed strand; "P219" and "P220" are shRNAs that include a 10 base pair 5' spacer sequence; "P227" and "P228" include an H1 promoter; and "P229" and "P230" are shRNAs that include a larger, 12 bp loop structure). FIG. 7A depicts levels of MHC class I surface expression in live cells (as assessed by mean fluorescent intensity (MFI)), for revised versions of the shRNA construct as indicated. FIG. 7B depicts quantitated MHC class I knockdown results at day 7 assessed by qRT-PCR, for the revised shRNA constructs indicated. In sum, strong knockdown (approx.70%) and reduced surface MHC class I levels (as assessed by MFI measurement) were observed with the "P220" revised shRNA construct. FIG. 8 depicts flow cytometry analyses that demonstrated observed rates of MHC class I knockdown at D3 and D7, when cells were administered no inhibitory RNA (top plots of MHC class I levels versus side scatter area (SSC-A)), a beta 2 microglobulin (B2M)-targeting siRNA as a positive control for knockdown, and the beta 2 microglobulin (B2M)-targeting shRNA in the test P220 shRNA expression construct. The P220 shRNA construct was observed to effectively and significantly reduce MHC class I expression levels, with peak shRNA-mediated knockdown of MHC class 1 observed at day 7. FIG.9 shows a bar graph of the dramatic reduction of MHC class I levels observed between day three (D3) and day seven (D7) when the P220 shRNA construct was administered. FIG.10 shows that the P220 shRNA construct was also effective at knocking down MHC class I levels in Jurkat cells at day 7 and day 11 post-administration, as compared to all other shRNA constructs tested. FIGs. 11A to 11C show that robust shRNA donor DNA design demonstrated greater than 60% knockdown of two different complexes (MHC class I and TCR) respectively targeted by shRNAs for beta 2 microglobulin (B2M) and CD3ζ, with such effects observed across different tested cell lines in vitro. FIG.11A shows that contacting of HH cells, Jurkat cells, or T cells with a B2M-targeting shRNA resulted in significant (>60% to 80%) knockdown of MHC class I in each tested cell line. Such results were consistent with previous data that demonstrated 70-75% knockdown of MHC class I. Effective knockdown of MHC class I is notably believed to be important for achieving persistence of a CAR-T-mediated effect (host vs. graft). FIG. 11B shows that contacting of Jurkat cells or T cells with a CD3ζ-targeting shRNA resulted in significant (>60% to 80%) knockdown of TCR complex in each tested cell line. Such results were consistent with previous data that demonstrated 60-80% knockdown of TCR complex. Effective knockdown of TCR complex is notably believed to be important for achieving safety of a CAR-T-mediated effect (by minimizing potential graft vs. host effects). FIG.11C shows cell sorting results that demonstrate the significant knockdown of TCR complex (and corresponding absence of antigen presenting cells) achieved by administration of CD3ζ-targeting shRNA to a population of T cells (bottom panel), as compared to an untreated control population of T cells (top panel). FIG.12 shows a schematic diagram of the P287 CD19-CAR B2M.2 shRNA P220 construct of the instant disclosure (SEQ ID NO: 1), having a 10 bp P220 linker and a 100 bp interpromoter spacer. The order and orientation of respective elongation factor 1 α (EF1a) – CD19 CAR and U6 promoter – shRNA sequences are shown at bottom. FIG.13 shows a schematic diagram of the P288 B2M.2 shRNA P220 CD19-CAR construct of the instant disclosure (SEQ ID NO: 2), having a 10 bp P220 linker and a 100 bp interpromoter spacer. The order and orientation of respective U6 promoter – shRNA and EF1a – CD19 CAR sequences are shown at bottom. FIG.14 shows a schematic diagram of the P289 B2M.2 shRNA P220 reverse CD19-CAR construct of the instant disclosure (SEQ ID NO: 3), having a 10 bp P220 linker and a 100 bp interpromoter spacer. The order and orientation of respective U6 promoter – shRNA and EF1a – CD19 CAR sequences are shown at bottom. FIGs. 15A and 15B show that the P287, P288 and P289 constructs of FIGs. 12-14 above achieved robust co-expression of both CD19-CAR and B2M-targeting shRNA, with B2M-targeting shRNA expression assessed via measurement of MHC class I knockdown in treated cells. FIG.15A shows that the P287, P288 and P289 constructs each expressed CD19-CAR at equal or greater levels than a CD19-CAR-only control ("P023") construct, in both Jurkat cells and T cells. FIG.15B shows that the B2M shRNA of each of the P287, P288 and P289 constructs was expressed and active, as demonstrated by robust (60-90%) levels of MHC class I knockdown observed for each such B2M- targeting shRNA-containing construct, in both Jurkat cells and T cells, as compared to the CD19- CAR-only negative control construct. FIG.16 shows an exemplary preparation schematic (manufacturing process) for allogeneic CAR cells of the instant disclosure, as well as (at lower left) demonstration by cell sorting of successful knock-in and expression of CAR (at approx. 90% levels) in cells treated with either a CD19-CAR only construct or with a CD19-CAR + B2M-targeting shRNA co-expression construct. At lower right, expression of the B2M-targeting shRNA of such co-expression construct, as observed by its efficacy in knocking down MHC class I at > 70% levels, was also demonstrated, as compared to the CD19-CAR only control construct. FIG.17 shows a schematic diagram of the P331 CD19-CAR CD3z shRNA1 construct of the instant disclosure (SEQ ID NO: 4), having an 11 bp P220 linker and a 100 bp interpromoter spacer. FIG.18 shows a schematic diagram of the P332 CD19-CAR CD3z shRNA2 construct of the instant disclosure (SEQ ID NO: 5), having an 11 bp P220 linker and a 100 bp interpromoter spacer. FIG.19 shows an exemplary expression cassette for multiplexed shRNAs (here, both B2M- targeting and CIITA-targeting shRNAs are present in a single construct). FIG.20 depicts an exemplary dual shRNA expression cassette for shRNAs that target CD3ζ and B2M, respectively. FIGs.21A to 21C show design of seventeen new shRNA base builds. FIG.21A depicts an exemplary parent P220 expression cassette and structure for a shRNA that targets B2M (top strand: SEQ ID NO: 78; bottom strand: SEQ ID NO: 79). FIG.21B shows a table of seventeen shRNA base builds, where design of five of the seventeen new shRNAs was based upon internal hits from HH and / or Jurkat cells (H1 promoter; Hit P228 – Target; Hits OL54 and OL64 – Reverse strands; Hit P218), while the remaining twelve shRNAs carried new modifications based upon the literature. FIG.21C depicts a two-dimensional schematic of an exemplary B2M-targeting shRNA cassette of the disclosure (SEQ ID NO: 80), showing respective spacer / leader, target (double-stranded region), loop and terminal domains. FIGs. 22A and 22B show that six of the seventeen new shRNA constructs screened in T cells showed similar knockdown as the parent P220 expression cassette shRNA (> 65% knockdown of MHC class I), while measurable benefits to knockdown levels were observed for shRNAs including new loop sequences and a 19A bulge. FIG. 22A depicts a bar graph demonstrating that the parent P220 B2M-targeting shRNA cassette produced an approximate 65% knockdown of MHC class I in T cells. Meanwhile, shRNA expression constructs showing equal or greater MHC class I knockdown in tested T cells than the P220 control included: a shRNA cassette having a 7 bp loop (70% knockdown); a shRNA cassette having a 9 bp loop (71% knockdown); a shRNA cassette having a 25 nt target (69% knockdown); a shRNA cassette having a 19A bulge (72% knockdown); a shRNA cassette having a downstream SV40 (66% knockdown); and a shRNA cassette having a downstream cPPT / CTS configuration (65% knockdown). FIG.22B depicts the results of cell sorting experiments, in which cells treated with the B2M shRNA were assessed for B2M shRNA-mediated knockdown of MHC class I. The tested B2M-targeting shRNA exhibited significant reduction of MHC class I levels, relative to an appropriate non-treated control population of cells. DETAILED DESCRIPTION OF THE INVENTION The present disclosure is based, at least in part, upon discovery and use of nucleic acid molecules that include CAR-encoding sequences and one or more shRNA-encoding sequences within a single nucleic acid molecule, with such molecules used to engineer cells for effective CAR- expressing cell-based therapies with enhanced safety and / or persistence attributes, as compared to CAR-expressing cells that do not also include shRNA-expressing sequences as a component of the nucleic acid molecule. In certain aspects, the CAR-encoding and shRNA-encoding sequences of the nucleic acid molecule are flanked by terminal repeats or other sequences capable of being recognized by an enzyme that can mediate genomic integration of the nucleic acid molecule. Introduction of the enzyme capable of performing targeted genomic integration, or of an additional nucleic acid (e.g., a DNA, a mRNA, etc.) encoding for and capable of expressing such an enzyme, to a cell in the presence of the nucleic acid molecule harboring CAR-encoding and shRNA-encoding sequences can be used to perform genomic integration of the nucleic acid molecule harboring CAR-encoding and shRNA-encoding sequences, thereby creating an engineered cell that can, in certain embodiments, be used as a cell-based CAR therapy, whether as an autologous cell therapy or as an allogeneic cell-based therapy (with shRNA-mediated knockdown of, e.g., MHC class I, MHC class II, and / or TCR molecules optionally reducing toxicity and / or extending persistence of such a cell- based allogenic therapy, as compared to a comparable CAR-expressing cell that does not include such shRNAs within the integrated, CAR-encoding nucleic acid molecule). Anti-CD19 CAR T cells, mainly generated from autologous peripheral blood lymphocytes, have shown remarkable clinical responses in patients with B cell-derived hematologic malignancies (June CH, Sadelain M. N Engl J Med. (2018) 379:64–73). Therapies based on autologous T cells confront severe limitations, mainly related to the fact that the product has to be generated from each patient's cells, in a time-consuming and costly process, and with the risk of manufacturing failure (Zhao et al. J Hematol Oncol. (2018) 11:132). Indeed, delay in treatment availability can be particularly problematic in patients with highly proliferative diseases (Depil et al. Nat Rev Drug Discov. (2020) 19:185–99). An additional hurdle lies in the quantity and quality of the starting autologous T cells as patients usually receive lymphodepleting chemotherapy and / or radiotherapy (Ceppi et al. Transfusion. (2018) 58:1414–20). In addition, heterogeneity of tumor antigen expression and immune evasion mechanisms developed by tumor cells require using CAR T cell products able to target multiple antigen specificities (Walsh et al. Curr Hematol Malig Rep. (2019) 14:451–9). In contrast, using T cells obtained from healthy donors (allogeneic T cells), can provide high amounts of fully functional cells and can allow for generation of multiple “off-the-shelf” CAR T cell products (Zhao et al.; Depil et al.). Successful therapeutic use of allogeneic CAR-T cells also confront certain challenges. Indeed, allogeneic T cells could cause severe graft-vs.-host disease (GVHD) and the host immune system could in turn induce allorejection, which will impede anti-tumor activity. There are different ways to avoid GVHD when designing allogeneic CAR T cells, the most widely used strategy being the generation of TCR-deficient T cells using genome editing tools such as Zinc finger nucleases (ZFN) (Provasi et al. Nat Med. (2012) 18:807–15; Torikai et al. Blood. (2012) 119:5697–705; Torikai et al. Blood. (2013) 122:1341–9), transcription activator-like effector nucleases (TALEN) (Poirot et al. Cancer Res. (2015) 75:3853–64; Rasaiyaah et al. JCI insight. (2018) 3:e99442; Sommer et al. Mol Ther. (2019) 27:1126–38) and CRISPR / Cas9 (Ren et al. Clin Cancer Res. (2017) 23:2255– 66; Eyquem et al. Nature. (2017) 543:113–7; Georgiadis et al. Mol Ther. (2018) 26:1215–27). Genetically eliminating key molecules governing CAR T cell immunogenicity is an appealing means of reducing allorejection, and disrupting the β2-microglobulin locus has been previously described (Wang et al. Stem Cells Transl Med. (2015) 4:1234–45). However, approaches that rely upon introduction of genomic disruptions in allogeneic cells present additional issues related to off-target integration events, particularly when needing to disrupt both loci of a target gene to be disrupted. Design and use of the shRNA-encoding / expressing nucleic acid molecules and engineered cells of the instant disclosure therefore provides an alternative to such knockout approaches, which is expected to improve therapeutic potential and reduce potentially toxic impacts of allogeneic cell therapies designed using the CAR / shRNA-expressing nucleic acid molecules of the instant disclosure. The following sections provide additional detail regarding elements and features of the nucleic acid molecules, engineered cells, and therapies of the instant disclosure, without limitation. Chimeric Antigen Receptors (CARs) CAR-T cells can act to kill targeted tumor cells, but can also elicit negative effects capable of propagating in an unchecked manner. Conventional CAR-T design uses a single polypeptide that includes tumor antigen-binding domains (e.g., an scFv), a transmembrane domain, a costimulatory domain and a CD3ζ domain (TCR ITAMs), with this single polypeptide therefore capable of specifically binding a tumor antigen and propagating an activated T cell response. (see, e.g., Wu et al. Science 350: aab4077.) As contemplated herein, the CARs (including CD-19-targeting CARs) of the present disclosure include an extracellular ligand binding domain capable of binding a targeted protein, typically an antigen, for example a tumor antigen. In one embodiment, the extracellular ligand binding domain is an antigen binding domain, for example, an antibody or an antigen binding fragment thereof. In particular embodiments, the antigen-binding fragment is a Fab or scFv. In one embodiment, the extracellular ligand binding domain is a ligand for a tumor marker, for example, a ligand that binds a marker expressed on the cell surface of a tumor, for example IL13 which binds to the IL13 receptor (IL13R) on glioma cells or heregulin which binds to erb B2, B3, and B4 on breast cancer cells. In one embodiment, the extracellular ligand binding domain targets a labeled or tagged protein or molecule, for example biotin or fluorescein isothiocyanate, which is bound to an antibody targeting a tumor expressed protein. For example, the extracellular ligand binding domain can target a label on a tumor-specific antibody, for example biotin, so that when the antibody-label binds to the tumor cell, the extracellular binding ligand of the CAR T-cell binds the label, activating the T-cell, and killing the tumor cell. In this regard, a “universal CAR” can be generated capable of binding any tagged or labeled antibody. See, e.g., Abate Daga et al.,“CAR models: next generation CAR modifications for enhanced T- cell function,” Molecular Therapy-Oncolytics (2016)3:1-7. In certain embodiments, the antigen binding domain in the CAR binds to a tumor antigen, for example, a tumor antigen associated with a hematological malignancy or a solid tumor. Tumor antigens capable of being targeted by CAR T-cells are known, and include, for example, but are not limited to, CD19, CD20, CD22, CD30, CD40, CD70, CD123, ErbB2 (HER2 / neu), epithelial cell adhesion molecule (EpCAM), Epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRvIII). Disialoganglioside GD2, disialoganglioside GD3, mesothelian, ROR1, mesothelin, CD33 / IL3Ra, C-Met, PSMA, Glycolipid, F77, EGFRvIII, GD-2, NY-ESO-1 TCR, melanoma-associated antigen (MAGE) A3 TCR, melanoma-associated antigen (MAGE) A1 TCR, alphafetapotein (AFP), carcinoembryonic antigen (CEA), CA-125, MUC-1, epithelial tumor antigen (ETA), tyrosinase, CA15-3, CA27-29, CA19-9, calcitonin, calretinin CD34, CD99MIC2, CD7, chromogranin, cytokeratin, desmin, CD31 FL1, glial fibrillary acidic protein, gross cystid disease fluid protein, HMB-45, human chorionic gonadotropin inhibin, MART-1, Myo D1, neuron- specific enolast, placental alkaline phosphatase, prostate specific antigens, PSCA. PTPRC, S100 protein, synaptophysin, thyroglobulin, thyroid transcription factor 1, tumor M2-PK, vimentin, human telomerase reverse transcriptase (hTERT), surviving, mouse double minute 2 homolog (MDM2), kappa-light chain, LeY, L1 cell adhesion molecule, oncofetal antigen (h5T4), TAG-72, VEGF-R2, and combinations thereof, as well as others described herein. Other antigens to which the antigen binding domain of the CAR can be directed include, but are not limited to, tissue or cell lineage specific antigens including, but not limited to, CD3, CD4, CD8, CD24, CD25, CD33, CD34, CD133, CD138, or a combination thereof. In certain embodiments, a CAR of the disclosure includes an extracellular domain having an antigen recognition domain, a transmembrane domain, and a cytoplasmic domain. In one embodiment, the transmembrane domain that naturally is associated with one of the domains in the CAR is used. In another embodiment, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex. In one embodiment, the transmembrane domain is the CD8α hinge domain. With respect to the cytoplasmic domain, the CAR of the disclosure can be designed to include at least one signaling domain and at least one co-stimulatory domain. Domains such as a FDA-approved drug-responsive heterodimer or a FDA-approved drug-inducible degron moiety may also be included. Heterodimer components and / or degron components of CARs have been disclosed in the art and are amino acid sequences to which, e.g., a FDA-approved drug can be bound, leading to activation in the presence of both drug and an antigen or to the degradation of the CAR when in contact, depending upon the CAR format used. As contemplated herein, the CARs of the present disclosure include a transmembrane domain spanning the extracellular ligand binding domain and the at least one intracellular co-stimulatory and / or signaling domain. Transmembrane domains useful in the construction of CARs are known in the art, and can be derived from natural or synthetic sources. For example, transmembrane regions contemplated herein include, but are not limited to, those derived from (i.e. comprise at least the transmembrane region(s) of) the alpha, beta or zeta chain of the T- cell receptor, CD28, CD3 epsilon, CD8, CD45, CD4, CD5, CDS, CD9, CD 16, CD22, CD33, CD37, CD64, CD80, CD86, CD 134, CD137, CD 154, or KIR2DS2. Alternatively, the transmembrane domain in some embodiments is synthetic, for example, comprising predominantly hydrophobic residues such as leucine and valine. In some aspects, a triplet of phenylalanine, tryptophan and valine will be found at each end of a synthetic transmembrane domain. As further contemplated herein, the CARs of the present disclosure include at least one intracellular (or cytoplasmic) signaling domain. The intracellular signaling domain of the CAR activates at least one of the normal effector functions or responses of the immune cell. For example, upon binding of the extracellular ligand domain to a target antigen, the signaling domain may act to activate the CAR T-cell, for example, by inducing a function of a T-cell such as cytolytic activity or T-helper activity, including the secretion of cytokines or other factors. In some embodiments, the CAR includes an intracellular component of the TCR complex, such as a TCR CD3+ chain that mediates T-cell activation and cytotoxicity, e.g., the immunoreceptor tyrosine-based activation motif (ITAM) domain CD3 zeta chain (CD3ζ). Thus, in some aspects as contemplated herein, the antigen binding molecule is linked to one or more cell signaling domains. In some embodiments, cell signaling domains include CD3 transmembrane domain, CD3 intracellular signaling domains, and / or other CD transmembrane domains. In some embodiments, the CAR further includes a portion of one or more additional molecules such as Fc receptor γ, for example FcεRIγ, CD8, CD4, CD25, or CD16. For example, in some aspects, the CAR includes a chimeric molecule between CD3-zeta (CD3ζ) or Fc receptor γ and CD8, CD4, CD25 or CD16. In one embodiment, the intracellular signaling domain is a Dap-12 derived signaling domain. The intracellular signaling domain, or cytoplasmic signaling domain, used interchangeably herein, of the CAR of the disclosure is responsible for activation of at least one of the normal effector functions of the immune cell in which the CAR has been placed. The term “effector function” refers to a specialized function of a cell. Effector function of a T-cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines. Thus, the term “intracellular signaling domain” refers to the portion of a protein which transduces the effector function signal and directs the cell to perform a specialized function. While usually the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal. The term intracellular signaling domain is thus meant to include any truncated portion of the intracellular signaling domain sufficient to transduce the effector function signal. Examples of intracellular signaling domains for use in the CAR of the disclosure include the cytoplasmic sequences of the T-cell receptor (TCR) and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any synthetic sequence that has the same functional capability. It is known that signals generated through the TCR alone may not be sufficient for full activation of the T-cell and that a secondary or co-stimulatory signal may also be required. Thus, T- cell activation can be said to be mediated by two distinct classes of cytoplasmic signaling sequence: those that initiate antigen-dependent primary activation through the TCR (primary cytoplasmic signaling sequences) and those that act in an antigen- independent manner to provide a secondary or co-stimulatory signal (secondary cytoplasmic signaling sequences). Primary cytoplasmic signaling sequences regulate primary activation of the TCR complex either in a stimulatory way, or in an inhibitory way. Primary cytoplasmic signaling sequences that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs or ITAMs. Examples of ITAM containing primary cytoplasmic signaling sequences that are of particular use in the disclosure include those derived from TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d. In one embodiment, the cytoplasmic signaling molecule in the CAR of the disclosure comprises a cytoplasmic signaling sequence derived from CD3 zeta. The cytoplasmic domain of the CAR can be designed to comprise the CD3-zeta signaling domain by itself or combined with any other desired cytoplasmic domain(s) useful in the context of the CAR of the disclosure. For example, the cytoplasmic domain of the CAR can comprise a CD3 zeta chain portion and a costimulatory signaling region. The costimulatory signaling region refers to a portion of the CAR comprising the intracellular domain of a costimulatory molecule. A costimulatory molecule is a cell surface molecule other than an antigen receptor or their ligands that is required for an efficient response of lymphocytes to an antigen. Examples of such molecules include CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function- associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and a ligand that specifically binds with CD83, and the like. Thus, any of the costimulatory elements known in the art as useful in the construction of CARs are within the scope of the disclosure. The cytoplasmic signaling sequences within the cytoplasmic signaling portion of the CAR of the disclosure may be linked to each other in a random or specified order. Optionally, a short oligo- or polypeptide linker, preferably between 2 and 10 amino acids in length may form the linkage. A glycine-serine doublet provides a particularly suitable linker. In one embodiment, the cytoplasmic domain is designed to comprise the signaling domain of CD3-zeta and the signaling domain of CD28. In another embodiment, the cytoplasmic domain is designed to comprise the signaling domain of CD3-zeta and the signaling domain of 4-1BB. In yet another embodiment, the cytoplasmic domain is designed to comprise the signaling domain of CD3- zeta and the signaling domain of CD28 and 4-1BB. In some embodiments, the intracellular signaling domain comprises a chimeric CD28 and OX40 co-stimulatory domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD27 co-stimulatory domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD27 and DAP10 co- stimulatory domain. In some embodiments, the intracellular signaling domain or domains include the cytoplasmic sequences of the T-cell receptor (TCR), and in some aspects also those of co-receptors that in the natural context act in concert with such receptor to initiate signal transduction following antigen receptor engagement, and / or any derivative or variant of such molecules, and / or any synthetic sequence that has the same functional capability. In the context of a natural TCR, full activation generally requires not only signaling through the TCR, but also a costimulatory signal. Thus, in some embodiments, to promote full activation, a component for generating secondary or co- stimulatory signal is also included in the CAR. In other embodiments, the CAR does not include a component for generating a costimulatory signal. In some aspects, an additional CAR is expressed in the same cell and provides the component for generating the secondary or costimulatory signal. In some aspects, the cell comprises a first CAR which contains signaling domains to induce the primary signal and a second CAR which binds to a second antigen and contains the component for generating a costimulatory signal. For example, a first CAR can be an activating CAR and the second CAR can be a costimulatory CAR. In some aspects, both CARs must be ligated in order to induce a particular effector function in the cell, which can provide specificity and selectivity for the cell type being targeted. In one embodiment, the cell comprises a first CAR which contains signaling domains to induce the primary signal and a costimulatory ligand molecule to stimulate other immune cells. See, e.g., Abate Daga et al.,“CAR models: next generation CAR modifications for enhanced T-cell function,” Molecular Therapy-Oncolytics (2016)3:1- 7. In some embodiments, the CAR includes a signaling domain and / or transmembrane portion of a costimulatory receptor, such as CD28, 4-1BB, OX40, DAP10, and ICOS. In some aspects, the same CAR includes both the activating and costimulatory components; in other aspects, the activating domain is provided by one polypeptide (e.g., a CAR) whereas the costimulatory component is provided by another polypeptide (e.g., a CAR or ligand recognizing another antigen). In certain embodiments, the intracellular signaling domain comprises a CD28 transmembrane and signaling domain linked to a CD3 (e.g., CD3-zeta) intracellular domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD28 and CD 137 (4- 1BB, TNFRSF9) co-stimulatory domain, linked to a CD3 zeta intracellular domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD28 or CD 137 (4- 1BB, TNFRSF9) co- stimulatory domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD28 and OX40 co-stimulatory domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD27 co-stimulatory domain. In some embodiments, the intracellular signaling domain comprises a chimeric CD27 and DAP10 co-stimulatory domain. In some embodiments, the CAR encompasses two or more costimulatory domain combined with an activation domain, e.g., primary activation domain, in the cytoplasmic portion. One example is a receptor including intracellular components of CD3-zeta, CD28, and 4-1BB. Other examples include a receptor including intracellular components of CD3- zeta, CD28, and OX40. In embodiments, the CAR comprises an extracellular domain that binds an antigen selected from CD19, CD20, CD22, CD30, CD33, CD44, CD44v6, CD44v7 / 8, CD70, CD79a, CD79b, CD123, CD138, CD171, CEA, CSPG4, EGFR, EGFR family including ErbB2 (HER2), EGFRvlll, EGP2, EGP40, EPCAM, EphA2, EpCAM, FAP, fetal AchR, GD2, GD3, Glypican-3 (GPC3), HLA- A1+MAGE1, HLAA2+MAGE1, HLA-A3+MAGE1, HLA-A1+NY-ESO-1, HLA-A2+NY-ESO-1, HLA-A3+NY-ESO-1, IL-11Rα, IL-13Rα2, Lambda, Lewis-Y, Kappa, Mesothelin, Muc1, Muc16, NCAM, NKG2D Ligands, NY-ESO-1, PRAME, PSCA, PSMA, ROR1, SSX, Survivin, TAG72, TEMs, VEGFR2, alpha folate receptor, 5T4, αvβ6 integrin, BCMA, B7-H3, B7-H6, and CAIX. In embodiments, the CAR comprises a transmembrane domain derived from a polypeptide selected from CD8α; CD4, CD28, CD45, PD-1, and CD152. In embodiments, the CAR comprises one or more intracellular costimulatory signaling domains selected from CD28, CD54 (ICAM), CD134 (OX40), CD137 (41BB), IL-2Rβ, CD152 (CTLA4), CD273 (PD-L2), CD274 (PD-L 1), and CD278 (ICOS). In embodiments, the CAR comprises a CD3 zeta (CD3ζ) signaling domain. CAR constructs may also use any of an array of art-recognized linkers (e.g., linker sequences, linker compounds, etc., e.g., as described in WO 2017 / 024318). shRNAs Any suitable molecule present in the engineered cell can be targeted by the instant shRNAs, noting that design and use of other RNA interference molecules (e.g., siRNAs, antisense oligonucleotides, etc.) is also contemplated. Typical examples of envisaged targets are: a MHC class I gene (e.g., B2M, TAP1, TAP2), a MHC class II gene (e.g., CIITA), a MHC coreceptor gene (e.g., HLA-F, HLA-G), a TCR chain (e.g., TRAC / TCRα chain), a CD3 chain (e.g., CD3ζ), NKBBiL, LTA, TNF, LTB, LST1, NCR3, AIF1, LY6, a heat shock protein (e.g. FIS PAIL, HSPA1A, HSPA1B), complement cascade, regulatory receptors (e.g. NOTCH4), HLA-DM, H LA-DO, RING1, CD52, CD247, HCP5, DGKA, DGKZ, MICA, MICB, ULBP1, ULBP2, ULBP3, ULBP4, ULBP5, ULBP6, 2B4, A2AR, BAX, BLIMP1, C160 (POLR3A) , CBL-B, CCR6, CD7, CD95, CD123, DGK [DGKA, DGKB, DGKD, DGKE, DKGG, DGKH, DGKI, DGKK, DGKQ, DGKZ], DNMT3A, DR4, DR5, EGR2, FABP4, FABP5, FASN, GMCSF, HPK1, IL-10R [IL10RA, IL10RB], IL2, LFA1, NEAT 1, NFkB (including RELA, RELB, NFkB2, NFkBl, REL), NKG2A, NR4A (including NR4A1, NR4A2, NR4A3), PD1, PI3KCD, PPP2RD2, SHIP1, SOAT1 , SOCS1, T-BET, TET2, TGFBR1, TGFBR2, TGFBR3, TIG IT, TIM3, TOX, and ZFP36L2. The successful expression of shRNA has largely been dependent upon coupling with a polymerase III (Pol III) promoter (e.g. HI, U6) that generate RNA species lacking a 5' cap and 3' polyadenylation, enabling processing of the shRNA duplex. Once transcribed, the shRNA undergoes processing, export from the nucleus, further processing and loading into the RNA-induced silencing complex (RISC) complex leading to the targeting degradation of mRNA of choice (Moore et al., 2010). Both the active sequence and the passenger sequence of each of the artificial shRNA nucleotide sequences are typically between 18 and 40 nucleotides long, more particularly between 18 and 30 nucleotides, most particularly between 19 and 25 nucleotides long. Cells containing at least two RNA interference molecules can have advantages, particularly therapeutic benefits. RNA interference molecules can indeed be directed against targets of which (over)expression is undesirable. Certain cells disclosed herein contain multiplexed RNA interference (e.g., shRNA) molecules. These can be directed against one or more targets which need to be downregulated (either targets within the cell, or outside of the cell if the shRNA is secreted). Each RNA interference molecule can target a different molecule, they can target the same molecule, or a combination thereof (i.e. more than one RNA molecule directed against one target, while only one RNA interference molecule is directed against a different target). When the RNA interference molecules are directed against the same target, they can target the same region, or they can target a different region. In other words, the RNA interference molecules can be identical or not when directed against the same target. In certain aspects, a CAR of the disclosure is directed against a tumor, and one or more RNA interference molecules (e.g., shRNAs) may enable and / or enhance CAR activity, and / or may interfere with tumor function, e.g. by targeting an immune checkpoint, directly downregulating a tumor target, targeting the tumor microenvironment. Alternatively or additionally, one or more of the RNA interference molecules may prolong persistence of the therapeutic cells, or otherwise alter a physiological response (e.g. interfering with GvHD or host versus graft reaction). CAR-shRNA Nucleic Acid Molecules / Constructs Some aspects of the instant disclosure provide nucleic acid molecules containing CAR- encoding sequences and shRNA-encoding sequences respectively expressed via appropriate promoter elements and formatted for robust CAR and / or shRNA expression, optionally flanked by terminal repeats capable of allowing genomic integration to occur in the presence of a terminal repeat-recognizing enzyme. Specific, non-limiting examples of nucleic acid constructs, as well as construct elements, include the following. (1) P287 CD19-CAR B2M.2 shRNA Construct (FIG.12) tcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggacgtcattgtcgatctccccatgcgagagtagggaactgcca ggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcctgagtaggacaaatccgc cgggagcggatttgaacgttgcgaagcaacggcccggagggtggcgggcaggacgcccgccataaactgccaggcatcaaattaagcag aaggccatcctgacggatggcctttttgcgtttctacaaactctctctggctaactagagaacccactgcttactggcttatcgaaatggtaccgc ggccgcttaacacttggattgcgggaaacgagttaagtcggctcgcgtgaattgcgcgtactccgcgggagccgtcttaactcggttcatatag atttgcggtggagtgcgggaaacgtgtaaactcgggccgattgtaactgcgtattaccaaatatttgttgtcgacgaattcggctccggtgcccg tcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcg gggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacg ttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgc cttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaagga gccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgct ttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacact ggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggcca ccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggtctcgcgccgccgtgtatcgccccgccctgggcggcaa ggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatggaggacgcggcgc tcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccggg cgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttccccacactg agtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaagcctc agacagtggttcaaagtttttttcttccatttcaggtgtcgtgagccaccatggccttaccagtgaccgccttgctcctgccgctggccttgctgctc cacgccgccaggccggacatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggcaag tcaggacatctctaagtatttgaattggtatcagcagaaaccagatggaactgttaaactcctgatctaccatacatcaagattacactcaggagtc ccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccacttacttttgccaacag ggtaatacgcttccgtacacgttcggaggggggactaagttggaaataacaggctccacctctggatccggcaagcccggatctggcgagg gatccaccaagggcgaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtcacatgcactgtctcagg ggtctcattacccgactatggtgtaagctggattcgccagcctccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccac atactataattcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttcttaaaaatgaacagtctgcaaactgatga cacagccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggggtcaaggaacctcagtcaccgtctcctcagc tgccgcagcgaagcccaccacgacgccagcgccgcgaccaccaacaccggcgcccaccatcgcgtcgcagcccctgtccctgcgccca gaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccttggccgg gacttgtggggtccttctcctgtcactggttatcaccctttactgcaaacggggcagaaagaaactcctgtatatattcaaacaaccatttatgaga ccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcagca ggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttgg acaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaaga taagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtac agccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctgatcctcaggtgcaggctgcctatcagaaggtggtggct ggtgtggccaatgccctggctcacaaataccactgagatctttttccctctgccaaaaattatggggacatcatgaagccccttgagcatctgact tctggctaataaaggaaatttattttcattgcaatagtgtgttggaattttttgtgtctctcactcggaaggacatatgggagggcaaatcatttaaaa catcagaatgagtatttggtttagagtttggcaacatatgcccatatgctggctgccatgaacaaaggttggctataaagaggtcatcagtatatg aaacagccccctgctgtccattccttattccatagaaaagccttgacttgaggttagattttttttatattttgttttgtgttatttttttctttaacatcccta aaattttccttacatgttttactagccagatttttcctcctctcctgactactcccagtcatagctgtccctcttctcttatggagatcagagcgcacgg agatcaccgagcgatctatcagatcggcgaccatttgtgaggtactggagccgagaggtaactacgatgccgctaagaacctctcggtcgag ggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatttgactgtaaacacaaagatattagta caaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagt atttcgatttcttggctttatatatcttgtggaaaggacgaaacaccggggggagcagagaattctcttattcaagagataagagaattctctgctc cctttttgcatgcactagtaattatttatgtactgaatagataaaaaaatgtctgtgattgaataaattttcattttttacacaagaaaccgaaaatttcat ijĵ ttcaatcgaacccatacttcaaaagatataggcattttaaactaactctgattttgcgcgggaaacctaaataattgcccgcgccatcttatattttgg cgggaaattcacccgacaccgtagtgttaagctagcgaagagcctagctacctactcgagtctagagggcccgtttaaacccgctgatcagcc tcgactctagcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactgga tctatcaacaggagtccaagctcagatcattgtcacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggc gtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagatacc aggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtg gcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccga ccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagc agagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgct gaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcaga ttacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttg gtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaagcccaatctgaataatgttacaaccaattaac caattctgattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaat gaaggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaacctatta atttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagtttatgcatttctttcca gacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcgcctgagcgaggcgaa atacgcgatcgctgttaaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatcaacaatattttcacctg aatcaggatattcttctaatacctggaatgctgtttttccggggatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgat ggtcggaagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaa ctctggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcatc catgttggaatttaatcgcggcctcgacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcat gatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacgggccagagctgca (SEQ ID NO: 1) (2) P289 B2M.2 shRNA CD19-CAR Construct (FIG.13) tcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggacgtcattgtcgatctccccatgcgagagtagggaactgcca ggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcctgagtaggacaaatccgc cgggagcggatttgaacgttgcgaagcaacggcccggagggtggcgggcaggacgcccgccataaactgccaggcatcaaattaagcag aaggccatcctgacggatggcctttttgcgtttctacaaactctctctggctaactagagaacccactgcttactggcttatcgaaatggtaccgc ggccgcttaacacttggattgcgggaaacgagttaagtcggctcgcgtgaattgcgcgtactccgcgggagccgtcttaactcggttcatatag atttgcggtggagtgcgggaaacgtgtaaactcgggccgattgtaactgcgtattaccaaatatttgttgtcgacgaattcgagggcctatttccc atgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatttgactgtaaacacaaagatattagtacaaaatacgtg acgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagtatttcgatttctt ggctttatatatcttgtggaaaggacgaaacaccggggggagcagagaattctcttattcaagagataagagaattctctgctccctttttagagc gcacggagatcaccgagcgatctatcagatcggcgaccatttgtgaggtactggagccgagaggtaactacgatgccgctaagaacctctcg gtcggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgc ctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtg cagtagtcgccgtgaacgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacg ggttatggcccttgcgtgccttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcg aggccttgcgcttaaggagccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcacctt cgcgcctgtctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgg gccaagatctgcacactggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcgggg cctgcgagcgcggccaccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggtctcgcgccgccgtgtatcgcc ccgccctgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaa atggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtga ctccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcga tggagtttccccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatctt ggttcattctcaagcctcagacagtggttcaaagtttttttcttccatttcaggtgtcgtgagccaccatggccttaccagtgaccgccttgctcctg ccgctggccttgctgctccacgccgccaggccggacatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcacc atcagttgcagggcaagtcaggacatctctaagtatttgaattggtatcagcagaaaccagatggaactgttaaactcctgatctaccatacatca agattacactcaggagtcccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgc cacttacttttgccaacagggtaatacgcttccgtacacgttcggaggggggactaagttggaaataacaggctccacctctggatccggcaag cccggatctggcgagggatccaccaagggcgaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcctccacgaaagggtctggagtggctgggagtaatat ggggtagtgaaaccacatactataattcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttcttaaaaatgaac agtctgcaaactgatgacacagccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggggtcaaggaacctca gtcaccgtctcctcagctgccgcagcgaagcccaccacgacgccagcgccgcgaccaccaacaccggcgcccaccatcgcgtcgcagcc cctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctg ggcgcccttggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaaacggggcagaaagaaactcctgtatatattcaa acaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactga gagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagag gagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaat ijķ gaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggccttta ccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctgatcctcaggtgcaggctgcctat cagaaggtggtggctggtgtggccaatgccctggctcacaaataccactgagatctttttccctctgccaaaaattatggggacatcatgaagcc ccttgagcatctgacttctggctaataaaggaaatttattttcattgcaatagtgtgttggaattttttgtgtctctcactcggaaggacatatgggag ggcaaatcatttaaaacatcagaatgagtatttggtttagagtttggcaacatatgcccatatgctggctgccatgaacaaaggttggctataaag aggtcatcagtatatgaaacagccccctgctgtccattccttattccatagaaaagccttgacttgaggttagattttttttatattttgttttgtgttatttt tttctttaacatccctaaaattttccttacatgttttactagccagatttttcctcctctcctgactactcccagtcatagctgtccctcttctcttatggag atcgcatgcactagtaattatttatgtactgaatagataaaaaaatgtctgtgattgaataaattttcattttttacacaagaaaccgaaaatttcatttc aatcgaacccatacttcaaaagatataggcattttaaactaactctgattttgcgcgggaaacctaaataattgcccgcgccatcttatattttggcg ggaaattcacccgacaccgtagtgttaagctagcgaagagcctagctacctactcgagtctagagggcccgtttaaacccgctgatcagcctc gactctagcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggatc tatcaacaggagtccaagctcagatcattgtcacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcg tttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagatacca ggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtgg cgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgac cgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagca gagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctg aagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagatt acgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggt catgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaagcccaatctgaataatgttacaaccaattaacca attctgattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatga aggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaacctattaatt tcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagtttatgcatttctttccaga cttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcgcctgagcgaggcgaaat acgcgatcgctgttaaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatcaacaatattttcacctgaa tcaggatattcttctaatacctggaatgctgtttttccggggatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgatgg tcggaagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaactc tggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcatccat gttggaatttaatcgcggcctcgacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgat gatatatttttatcttgtgcaatgtaacatcagagattttgagacacgggccagagctgca (SEQ ID NO: 2) (3) P289 B2M.2 shRNA reverse CD19-CAR Construct (FIG.14) tcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggacgtcattgtcgatctccccatgcgagagtagggaactgcca ggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcctgagtaggacaaatccgc cgggagcggatttgaacgttgcgaagcaacggcccggagggtggcgggcaggacgcccgccataaactgccaggcatcaaattaagcag aaggccatcctgacggatggcctttttgcgtttctacaaactctctctggctaactagagaacccactgcttactggcttatcgaaatggtaccgc ggccgcttaacacttggattgcgggaaacgagttaagtcggctcgcgtgaattgcgcgtactccgcgggagccgtcttaactcggttcatatag atttgcggtggagtgcgggaaacgtgtaaactcgggccgattgtaactgcgtattaccaaatatttgttgtcgacgaattcaaaaagggagcag agaattctcttatctcttgaataagagaattctctgctccccccggtgtttcgtcctttccacaagatatataaagccaagaaatcgaaatactttcaa gttacggtaagcatatgatagtccattttaaaacataattttaaaactgcaaactacccaagaaattattactttctacgtcacgtattttgtactaatat ctttgtgtttacagtcaaattaattccaattatctctctaacagccttgtatcgtatatgcaaatatgaaggaatcatgggaaataggccctcagagc gcacggagatcaccgagcgatctatcagatcggcgaccatttgtgaggtactggagccgagaggtaactacgatgccgctaagaacctctcg gtcggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgc ctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtg cagtagtcgccgtgaacgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacg ggttatggcccttgcgtgccttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcg aggccttgcgcttaaggagccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcacctt cgcgcctgtctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgg gccaagatctgcacactggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcgggg cctgcgagcgcggccaccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggtctcgcgccgccgtgtatcgcc ccgccctgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaa atggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtga ctccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcga tggagtttccccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatctt ggttcattctcaagcctcagacagtggttcaaagtttttttcttccatttcaggtgtcgtgagccaccatggccttaccagtgaccgccttgctcctg ccgctggccttgctgctccacgccgccaggccggacatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcacc atcagttgcagggcaagtcaggacatctctaagtatttgaattggtatcagcagaaaccagatggaactgttaaactcctgatctaccatacatca agattacactcaggagtcccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgc cacttacttttgccaacagggtaatacgcttccgtacacgttcggaggggggactaagttggaaataacaggctccacctctggatccggcaag cccggatctggcgagggatccaccaagggcgaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcctccacgaaagggtctggagtggctgggagtaatat ggggtagtgaaaccacatactataattcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttcttaaaaatgaac agtctgcaaactgatgacacagccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggggtcaaggaacctca gtcaccgtctcctcagctgccgcagcgaagcccaccacgacgccagcgccgcgaccaccaacaccggcgcccaccatcgcgtcgcagcc cctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctg ggcgcccttggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaaacggggcagaaagaaactcctgtatatattcaa acaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactga gagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagag gagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaat gaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggccttta ccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctgatcctcaggtgcaggctgcctat cagaaggtggtggctggtgtggccaatgccctggctcacaaataccactgagatctttttccctctgccaaaaattatggggacatcatgaagcc ccttgagcatctgacttctggctaataaaggaaatttattttcattgcaatagtgtgttggaattttttgtgtctctcactcggaaggacatatgggag ggcaaatcatttaaaacatcagaatgagtatttggtttagagtttggcaacatatgcccatatgctggctgccatgaacaaaggttggctataaag aggtcatcagtatatgaaacagccccctgctgtccattccttattccatagaaaagccttgacttgaggttagattttttttatattttgttttgtgttatttt tttctttaacatccctaaaattttccttacatgttttactagccagatttttcctcctctcctgactactcccagtcatagctgtccctcttctcttatggag atcgcatgcactagtaattatttatgtactgaatagataaaaaaatgtctgtgattgaataaattttcattttttacacaagaaaccgaaaatttcatttc aatcgaacccatacttcaaaagatataggcattttaaactaactctgattttgcgcgggaaacctaaataattgcccgcgccatcttatattttggcg ggaaattcacccgacaccgtagtgttaagctagcgaagagcctagctacctactcgagtctagagggcccgtttaaacccgctgatcagcctc gactctagcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggatc tatcaacaggagtccaagctcagatcattgtcacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcg tttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagatacca ggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtgg cgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgac cgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagca gagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctg aagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagatt acgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggt catgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaagcccaatctgaataatgttacaaccaattaacca attctgattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatga aggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaacctattaatt tcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagtttatgcatttctttccaga cttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcgcctgagcgaggcgaaat acgcgatcgctgttaaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatcaacaatattttcacctgaa tcaggatattcttctaatacctggaatgctgtttttccggggatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgatgg tcggaagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaactc tggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcatccat gttggaatttaatcgcggcctcgacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgat gatatatttttatcttgtgcaatgtaacatcagagattttgagacacgggccagagctgca (SEQ ID NO: 3) (4) P331 CD19-CAR CD3z.1 shRNA Construct (FIG.17) tcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggacgtcattgtcgatctccccatgcgagagtagggaactgcca ggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcctgagtaggacaaatccgc cgggagcggatttgaacgttgcgaagcaacggcccggagggtggcgggcaggacgcccgccataaactgccaggcatcaaattaagcag aaggccatcctgacggatggcctttttgcgtttctacaaactctctctggctaactagagaacccactgcttactggcttatcgaaatggtaccgc ggccgcttaacacttggattgcgggaaacgagttaagtcggctcgcgtgaattgcgcgtactccgcgggagccgtcttaactcggttcatatag atttgcggtggagtgcgggaaacgtgtaaactcgggccgattgtaactgcgtattaccaaatatttgttgtcgacgaattcggctccggtgcccg tcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcg gggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacg ttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgc cttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaagga gccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgct ttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacact ggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggcca ccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggtctcgcgccgccgtgtatcgccccgccctgggcggcaa ggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatggaggacgcggcgc tcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccggg cgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttccccacactg agtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaagcctc agacagtggttcaaagtttttttcttccatttcaggtgtcgtgagccaccatggccttaccagtgaccgccttgctcctgccgctggccttgctgctc cacgccgccaggccggacatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggcaag tcaggacatctctaagtatttgaattggtatcagcagaaaccagatggaactgttaaactcctgatctaccatacatcaagattacactcaggagtc ccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccacttacttttgccaacag ggtaatacgcttccgtacacgttcggaggggggactaagttggaaataacaggctccacctctggatccggcaagcccggatctggcgagg gatccaccaagggcgaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtcacatgcactgtctcagg ggtctcattacccgactatggtgtaagctggattcgccagcctccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccac atactataattcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttcttaaaaatgaacagtctgcaaactgatga cacagccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggggtcaaggaacctcagtcaccgtctcctcagc tgccgcagcgaagcccaccacgacgccagcgccgcgaccaccaacaccggcgcccaccatcgcgtcgcagcccctgtccctgcgccca gaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccttggccgg gacttgtggggtccttctcctgtcactggttatcaccctttactgcaaacggggcagaaagaaactcctgtatatattcaaacaaccatttatgaga ccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcagca ggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttgg acaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaaga taagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtac agccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaatcctcaggtgcaggctgcctatcagaaggtggtggct ggtgtggccaatgccctggctcacaaataccactgagatctttttccctctgccaaaaattatggggacatcatgaagccccttgagcatctgact tctggctaataaaggaaatttattttcattgcaatagtgtgttggaattttttgtgtctctcactcggaaggacatatgggagggcaaatcatttaaaa catcagaatgagtatttggtttagagtttggcaacatatgcccatatgctggctgccatgaacaaaggttggctataaagaggtcatcagtatatg aaacagccccctgctgtccattccttattccatagaaaagccttgacttgaggttagattttttttatattttgttttgtgttatttttttctttaacatcccta aaattttccttacatgttttactagccagatttttcctcctctcctgactactcccagtcatagctgtccctcttctcttatggagatcagagcgcacgg agatcaccgagcgatctatcagatcggcgaccatttgtgaggtactggagccgagaggtaactacgatgccgctaagaacctctcggtcgag ggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatttgactgtaaacacaaagatattagta caaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagt atttcgatttcttggctttatatatcttgtggaaaggacgaaacaccgggtgccttgttcctgagagtgaaatattcattcactctcaggaacaaggc atttttgcatgcactagtaattatttatgtactgaatagataaaaaaatgtctgtgattgaataaattttcattttttacacaagaaaccgaaaatttcattt caatcgaacccatacttcaaaagatataggcattttaaactaactctgattttgcgcgggaaacctaaataattgcccgcgccatcttatattttggc gggaaattcacccgacaccgtagtgttaagctagcgaagagcctagctacctactcgagtctagagggcccgtttaaacccgctgatcagcct cgactctagcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggat ctatcaacaggagtccaagctcagatcattgtcacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggc gtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagatacc aggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtg gcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccga ccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagc agagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgct gaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcaga ttacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttg gtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaagcccaatctgaataatgttacaaccaattaac caattctgattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaat gaaggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaacctatta atttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagtttatgcatttctttcca gacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcgcctgagcgaggcgaa atacgcgatcgctgttaaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatcaacaatattttcacctg aatcaggatattcttctaatacctggaatgctgtttttccggggatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgat ggtcggaagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaa ctctggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcatc catgttggaatttaatcgcggcctcgacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcat gatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacgggccagagctgca (SEQ ID NO: 4) (5) P332 CD19-CAR CD3z.2 shRNA Construct (FIG.18) tcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggacgtcattgtcgatctccccatgcgagagtagggaactgcca ggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcctgagtaggacaaatccgc cgggagcggatttgaacgttgcgaagcaacggcccggagggtggcgggcaggacgcccgccataaactgccaggcatcaaattaagcag aaggccatcctgacggatggcctttttgcgtttctacaaactctctctggctaactagagaacccactgcttactggcttatcgaaatggtaccgc ggccgcttaacacttggattgcgggaaacgagttaagtcggctcgcgtgaattgcgcgtactccgcgggagccgtcttaactcggttcatatag atttgcggtggagtgcgggaaacgtgtaaactcgggccgattgtaactgcgtattaccaaatatttgttgtcgacgaattcggctccggtgcccg tcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcg gggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacg ttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgc cttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaagga gccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgct ttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacact ggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggcca ccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggtctcgcgccgccgtgtatcgccccgccctgggcggcaa ggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatggaggacgcggcgc tcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccggg cgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttccccacactg agtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaagcctc agacagtggttcaaagtttttttcttccatttcaggtgtcgtgagccaccatggccttaccagtgaccgccttgctcctgccgctggccttgctgctc cacgccgccaggccggacatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggcaag tcaggacatctctaagtatttgaattggtatcagcagaaaccagatggaactgttaaactcctgatctaccatacatcaagattacactcaggagtc ccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccacttacttttgccaacag ggtaatacgcttccgtacacgttcggaggggggactaagttggaaataacaggctccacctctggatccggcaagcccggatctggcgagg gatccaccaagggcgaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtcacatgcactgtctcagg ggtctcattacccgactatggtgtaagctggattcgccagcctccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccac atactataattcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttcttaaaaatgaacagtctgcaaactgatga cacagccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggggtcaaggaacctcagtcaccgtctcctcagc tgccgcagcgaagcccaccacgacgccagcgccgcgaccaccaacaccggcgcccaccatcgcgtcgcagcccctgtccctgcgccca gaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccttggccgg gacttgtggggtccttctcctgtcactggttatcaccctttactgcaaacggggcagaaagaaactcctgtatatattcaaacaaccatttatgaga ccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcagca ggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttgg acaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaaga taagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtac agccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaatcctcaggtgcaggctgcctatcagaaggtggtggct ggtgtggccaatgccctggctcacaaataccactgagatctttttccctctgccaaaaattatggggacatcatgaagccccttgagcatctgact tctggctaataaaggaaatttattttcattgcaatagtgtgttggaattttttgtgtctctcactcggaaggacatatgggagggcaaatcatttaaaa catcagaatgagtatttggtttagagtttggcaacatatgcccatatgctggctgccatgaacaaaggttggctataaagaggtcatcagtatatg aaacagccccctgctgtccattccttattccatagaaaagccttgacttgaggttagattttttttatattttgttttgtgttatttttttctttaacatcccta aaattttccttacatgttttactagccagatttttcctcctctcctgactactcccagtcatagctgtccctcttctcttatggagatcagagcgcacgg agatcaccgagcgatctatcagatcggcgaccatttgtgaggtactggagccgagaggtaactacgatgccgctaagaacctctcggtcgag ggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatttgactgtaaacacaaagatattagta caaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaaagt atttcgatttcttggctttatatatcttgtggaaaggacgaaacaccgggacccgtcaatgtactaggataatattcatatcctagtacattgacggg ttttttgcatgcactagtaattatttatgtactgaatagataaaaaaatgtctgtgattgaataaattttcattttttacacaagaaaccgaaaatttcattt caatcgaacccatacttcaaaagatataggcattttaaactaactctgattttgcgcgggaaacctaaataattgcccgcgccatcttatattttggc gggaaattcacccgacaccgtagtgttaagctagcgaagagcctagctacctactcgagtctagagggcccgtttaaacccgctgatcagcct cgactctagcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggat ctatcaacaggagtccaagctcagatcattgtcacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggc gtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagatacc aggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtg gcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccga ccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagc agagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgct gaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcaga ttacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttg gtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaagcccaatctgaataatgttacaaccaattaac caattctgattagaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaat gaaggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaacctatta atttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagtttatgcatttctttcca gacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcgcctgagcgaggcgaa atacgcgatcgctgttaaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatcaacaatattttcacctg aatcaggatattcttctaatacctggaatgctgtttttccggggatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgat ggtcggaagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaa ctctggcgcatcgggcttcccatacaagcgatagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatcagcatc catgttggaatttaatcgcggcctcgacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaagcagacagttttattgttcat gatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacgggccagagctgca (SEQ ID NO: 5) Exemplary CAR amino acid sequences encoded for by exemplified or otherwise contemplated nucleic acid molecules of the instant disclosure include the following. CD19-CAR amino acid sequence (4-1BB / CD3z) MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKP DGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGT KLEITGSTSGSGKPGSGEGSTKGEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIR QPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKH YYYGGSYAMDYWGQGTSVTVSSAAAAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAG GAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQE EDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGR DPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKD TYDALHMQALPPR* (SEQ ID NO: 6) CD20-CAR amino acid sequence (4-1BB / CD3z) MLLLVTSLLLCELPHPAFLLIPEVQLQQSGAELVKPGASVKMSCKASGYTFTSYNMHWVK QTPGQGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSADYYCAR SNYYGSSYWFFDVWGAGTTVTVSSGGGGSGGGGSGGGGSDIVLTQSPAILSASPGEKVT MTCRASSSVNYMDWYQKKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAE DAATYYCQQWSFNPPTFGGGTKLEIKAAATTTPAPRPPTPAPTIASQPLSLRPEACRPAAG GAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQE EDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGR DPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKD TYDALHMQALPPR* (SEQ ID NO: 7) CD22-CAR amino acid sequence (4-1BB / CD3z) MLLLVTSLLLCELPHPAFLLIPQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIR QSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCA REVTGDLEDAFDIWGQGTMVTVSSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQTIWS YLNWYQQRPGKAPNLLIYAASSLQSGVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSY SIPQTFGQGTKLEIKAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFAC DIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEE GGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKN PQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPP R* (SEQ ID NO: 8) B7H3-CAR (MGA271; 4-1BB, CD3z) MALPVTALLLPLALLLHAARPEVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWVR QAPGKGLEWVAYISSDSSAIYYADTVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCG RGRENIYYGSRLDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIQLTQSPSFLSASVGDRVT ITCKASQNVDTNVAWYQQKPGKAPKALIYSASYRYSGVPSRFSGSGSGTDFTLTISSLQPE DFATYYCQQYNNYPFTFGQGTKLEIKAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGG AVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEE DGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRD PEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDT YDALHMQALPPR* (SEQ ID NO: 9) B7H3-CAR (BRCA84; CD28, CD3z) MALPVTALLLPLALLLHAARPDIAMTQSQKFMSTSVGDRVSVTCKASQNVDTNVAWYQ QKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTINNVQSEDLAEYFCQQYNNYPFTF GSGTKLEIKGGGGSGGGGSGGGGSDVQLVESGGGLVQPGGSRKLSCAASGFTFSSFGMH WVRQAPEKGLEWVAYISSDSSAIYYADTVKGRFTISRDNPKNTLFLQMTSLRSEDTAMYY CGRGRENIYYGSRLDYWGQGTTLTVSSIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFP GPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHY QPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPE MGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTY DALHMQALPPR* (SEQ ID NO: 10) Her2-CAR amino acid sequence (CD28 / CD3z) MDFQVQIFSFLLISASVIMSRGDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQK PGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGT KVEIKRTGSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQ APGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSR WGGDGFYAMDVWGQGTLVTVSSIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSK PFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYA PPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGK PRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHM QALPPR* (SEQ ID NO: 11) An exemplary U6 promoter, used to promote shRNA expression in certain nucleic acid molecules / constructs of the disclosure, is provided below. gagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattggaattaatttgactgtaaacacaaagatatta gtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaacttgaa agtatttcgatttcttggctttatatatcttgtggaaaggac (SEQ ID NO: 12) Exemplary shRNA-encoding sequences of the disclosure include the following. B2M-targeting shRNA cassette gaaacaccggggggagcagagaattctcttattcaagagataagagaattctctgctcccttttt (SEQ ID NO: 13) TRAC-targeting shRNA cassette gaaacaccggggcagtgcttccaagataatcttcaagagagattatcttggaagcactgcttttt (SEQ ID NO: 14) CD3ζ-targeting shRNA cassette gaaacaccgggacccgtcaatgtactaggatatcaagagtatcctagtacattgacgggtttttt (SEQ ID NO: 15) Certain validated RNAi (e.g., shRNA) target sequences are provided in Table 1 below. Table 1. Additional Validated RNAi Target Sequences SEQ ID Complex Gene Target Target sequence 5'-3' NO: CIITA GGCUACCUGGAGCUUCUUAAC 35 CIITA.2 GCUCAGGCUAAGCUUGUACAA 36 An exemplary dual-targeting B2M-CIITA shRNA construct element (dual shRNA cassette; FIG.19) of the disclosure is provided by the following. gaaacaccggggggagcagagaattctcttattcaagagataagagaattctctgctcccttcgagcatggcgctcaggctaagcttgtacaat caagagttgtacaagcttagcctgagcttttt (SEQ ID NO: 57) An exemplary dual-targeting CD3z-B2M shRNA construct element (dual shRNA cassette; FIG.20) of the disclosure is provided by the following. gaaacaccgggacccgtcaatgtactaggataatattcatatcctagtacattgacgggtcgagcatggcgggagcagagaattctcttaattca agagataagagaattctctgctcccttttt (SEQ ID NO: 58) Exemplary linker sequences used between shRNA-encoding sequences expressed by a single promoter (e.g., a U6 promoter as exemplified herein) of the instant disclosure include the following. 10 bp linker #1: CGAGCATGGC (SEQ ID NO: 59) 10 bp linker #2: GTAGCACCAG (SEQ ID NO: 60) 10 bp linker #3: CACGTGCATC (SEQ ID NO: 61) 20 bp linker: TCAGACTAGAGCGGGGCTGT (SEQ ID NO: 62) 40 bp linker: TCAACGTGCTCCACGGCAGGCGCCTGACGAGGGGCCCACA (SEQ ID NO: 63) 80 bp linker: CCGAGGAAGTAGACTGTTGCACGTTGGGGATAGCGGTAGCTAACTAAGACGC CTGCCACAACAGCAGTATCAAACCCGTA (SEQ ID NO: 64) 120 bp linker: AAGTTTTATCTAAACACGATGAGAAGAGTATTCGTCAGGCCACATGGCTATCT TGTTCTGGTCGGATCCATCGTTGGCGCCCGACCACCACATTCCATAGTGAGTTCTTCGT CCGAG CCA (SEQ ID NO: 65) Targeted Genomic Integration Enzymes Certain aspects of the disclosure feature the design and use of nucleic acid molecules having exogenous sequences (e.g., CAR-encoding, shRNA-encoding, etc.) that are flanked by sequences capable of being recognized by an enzyme that can perform targeted genomic integration of the nucleic acid molecule, e.g., into the genome of a cell, e.g., an immune cell used for therapeutic administration to a subject in need thereof. A recombinase recognizes and binds specific sequences at the ends of a transposon, mediates synaptic interactions between the ends to bring them together, interacts with the target DNA, and executes the DNA breakage and joining reactions that underlie recombination. An integrase is a recombinase enzyme that is capable of integrating DNA (e.g., of a virus), into another piece of DNA, usually the host chromosome. DNA transposons are mobile elements that use a “cut-and-paste” mechanism. DNA is excised by double strand cleavage from the donor molecule and integrated into the acceptor molecule. Transposons move from one position on DNA to a second position on DNA in the presence of a transposase, an enzyme that binds to the end of a transposon and catalyzes its movement to a specific genomic location in a host. A main concern in transposase-based gene therapy is insertional mutagenesis due to random integration, albeit mostly at known sequences (e.g. TTAA sequences), near or within loci that activate oncogenes, interrupt tumor-suppressor genes, or disrupt the transcription of normal genes. Thus, while non-viral, transposon gene therapy approaches have great promise for treating individuals with genetic disorders, the challenge is to reduce the risk of random insertion. Accordingly, there is a clinical need to improve the safety of enzymes capable of transposition, such as, e.g., transposases, to reduce the risk of insertional mutagenesis and oncogenesis. In embodiments, transpose enzymes with reduced risk of insertional mutagenesis and oncogenesis can be used in the nucleic acid molecules and therapeutic cells of the instant disclosure. In exemplary embodiments, the enzyme is derived from Bombyx mori, Xenopus tropicalis, Trichoplusiani, or Myotis lucifugus. In embodiments, the enzyme is an engineered version, including but not limited to an enzyme that is a monomer, dimer, tetramer (or another multimer), hyperactive, or has a reduced interaction with non-TTAA recognitions sites (Int−), derived from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, or Myotis lucifugus. In some embodiments, the transposase enzyme is a Myotis lucifugus transposase (referred to herein as MLT or an MLT transposase), which can be either the wild type, monomer, dimer, tetramer (or another multimer), hyperactive, an Int- mutant, or any other variant, e.g., as described in U.S. Patent No. 11,542,528, International Application No. PCT / US2022 / 079288, and / or U.S. Serial No.63 / 479,015. In embodiments, the transposase enzyme is derived from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Myotis lucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, troglodytes, Molossus molossus, or Homo sapiens. In embodiments, the transposase enzyme is derived from any of Trichoplusia ni, Myotis lucifugus, Myotis myotis, Pan troglodytes, or Pteropus vampyrus. The transposases can have one or more hyperactive and / or integration deficient mutations, or equivalents thereof. One skilled in the art can correspond such mutants to transposases from any of Trichoplusia ni, Myotis lucifugus, Myotis myotis, or Pteropus vampyrus. Also, one skilled in the art can correspond such mutants to transposases from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Myotis lucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, troglodytes, Molossus molossus, or Homo sapiens. In some embodiments, the enzyme (e.g., without limitation, a transposase) has a nucleotide sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity to a nucleotide sequence of any of Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, and Molossus molossus. In some embodiments, the transposase enzyme can have an amino acid sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity to an amino acid sequence of any of Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, and Molossus molossus. In some aspects, a nucleic acid encoding an enzyme capable of targeted genomic integration by transposition (e.g., a chimeric transposase) in accordance with embodiments of the present disclosure is provided. The nucleic acid can be DNA or RNA. In some embodiments, the nucleic acid encoding the enzyme is DNA. In some embodiments, the nucleic acid encoding the enzyme capable of targeted genomic integration by transposition (e.g., a chimeric transposase) is RNA such as, e.g., helper RNA. In embodiments, the chimeric transposase is incorporated into a vector. In some embodiments, the vector is a non-viral vector. In embodiments, a nucleic acid encoding a transposon is a DNA, referred to as a “donor DNA.” In embodiments, a nucleic acid encoding an enzyme capable of targeted genomic integration by transposition (e.g., a chimeric transposase) is helper RNA. In embodiments, the donor DNA is incorporated into a plasmid. In embodiments, the donor DNA is a plasmid. In some aspects, a host cell comprising the nucleic acid in accordance with embodiments of the present disclosure is provided. In certain embodiments, an enzyme capable of targeted genomic integration by transposition (e.g., a recombinase, an integrase, or a transposase enzyme), as a monomer or a dimer, can be fused with a transcription activator-like effector protein (TALE) DNA binding domain (DBD), a zinc finger (ZnF) nucleic acid binding domain, and / or a dCas9 / gRNA, to thereby create a chimeric enzyme capable of a site- or locus-specific transposition. Without wishing to be bound by theory, the enzyme (e.g., without limitation, a chimeric transposase) utilizes the specificity of TALE DBD and / or ZnF binding domains to certain sites within a host genome, which allows using DBDs to target any desired location in the genome with the nucleic acid molecule of the disclosure. In this way, the chimeric transposase in accordance with the present disclosure allows achieving targeted integration of a nucleic acid molecule disclosed herein. In embodiments, the enzyme capable of targeted genomic integration by transposition is a recombinase or an integrase. In embodiments, the recombinase is an integrase. In embodiments, the integrase is a transposase or the recombinase is a transposase. As discussed herein and known in the art, viral (e.g. AAV, lentivirus, etc.) and nuclease- based (e.g. CRISPR / Cas, prime editing base editing) gene therapies are typically limited by mutagenesis risk, and also have drawbacks such as immunogenicity, manufacturing costs, cargo size, and reversibility. Transposons are less likely to activate a proto-oncogene than lentivirus or other retroviruses but cause insertional mutagenesis when a transgene is inserted in one or more locations in a host genome other than the intended locations. In particular, the genomic sites recognized by transposases, such as a TA dinucleotide site or a TTAA tetranucleotide site, can be found in multiple locations in a genome such that a transgene can be inserted in unintended locations within the genome and have disruptive, often severe effects on the host. For example, the insertional mutagenesis can affect a function of a metabolic gene. Accordingly, to improve the function of a dual transposon / transposase system as a safe and efficient gene therapy tool, it is desired to increase and control the specificity of a transposase's binding and insertion. Accordingly, in some aspects, a composition is provided that comprises an enzyme capable of transposition, comprising (a) a TALE DBD or a dCas9 / gRNA complex; (b) an enzyme capable of targeted genomic integration by transposition, the enzyme being capable of inserting a transposon at a TA dinucleotide site or a TTAA tetranucleotide site in a GSHS in a nucleic acid molecule; and (c) a linker that connects the TALE DBD or the dCas9 / gRNA complex and the enzyme. In embodiments, the enzyme (e.g., a transposase enzyme) is a head to tail dimer. In some embodiments, the enzyme is a tetramer. In some embodiments, the enzyme is a monomer. In embodiments, an enzyme capable of targeted genomic integration by transposition is encoded by a first nucleic acid, and the transposon (e.g., nucleic acid molecule including CAR- encoding and shRNA-encoding sequences) is encoded by a second, non-viral nucleic acid. The transposon comprises, e.g., a nucleic acid molecule including CAR-encoding and shRNA-encoding sequences and is flanked by ends recognized by the enzyme, and the enzyme causes the transgene to be inserted in a certain genomic locus and / or site (e.g., at a TA dinucleotide site or a TTAA tetranucleotide site in a genomic safe harbor site (GSHS) of a nucleic acid molecule). In some embodiments, the first nucleic acid is RNA, for example, helper RNA; and the second, non-viral nucleic acid is DNA. In embodiments, inteins (also referred to as splicing domains) are used to synthesize a recombinant enzyme (e.g., without limitation, an MLT fusion protein) that includes desired internal DNA biding domains (DNA binders) that target specific sites within the human genome for integration of a donor transgene. In embodiments, an enzyme capable of targeted genomic integration by transposition is, without limitation, a transposase enzyme. In embodiments, the transposase enzyme is derived from Bombyx mori, Xenopus tropicalis, or Trichoplusia ni. In embodiments, the enzyme (e.g., without limitation, a transposase enzyme) is an engineered version of a transposase enzyme, including but not limited to monomers, dimers, tetramers, hyperactive, or Int-forms, derived from Bombyx mori, Xenopus tropicalis, or Trichoplusia ni. In embodiments, the transposase enzyme is an engineered version, including but not limited to a transposase enzyme that is a monomer, dimer, tetramer, hyperactive, or has a reduced interaction with non-TTAA recognitions sites (Int−), derived from any of Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Myotis lucifugus, Pipistrellus kuhlii, Pteropus vampyrus, and Molossus molossus Bombyx mori, Xenopus tropicalis, Trichoplusia ni or Myotis lucifugus. The transposase enzyme can be either the wild type, monomer, dimer, tetramer, hyperactive, or an Int-mutant. Any of a variety of terminal repeat sequences known in the art, as well as associated terminal repeat-recognizing enzymes capable of performing genomic integration, are contemplated for use in the nucleic acid molecules / constructs of the disclosure. The following are examples of such terminal repeat-recognizing enzymes, and of certain terminal repeat sequences. Myotis lucifugus mobile element enzyme (MLT) MAQHSDYSDDEFCADKLSNYSCDSDLENASTSDEDSSDDEVMVRPRTLRRRRISSSSSDSE SDIEGGREEWSHVDNPPVLEDFLGHQGLNTDAVINNIEDAVKLFIGDDFFEFLVEESNRYY NQNRNNFKLSKKSLKWKDITPQEMKKFLGLIVLMGQVRKDRRDDYWTTEPWTETPYFG KTMTRDRFRQIWKAWHFNNNADIVNESDRLCKVRPVLDYFVPKFINIYKPHQQLSLDEGI VPWRGRLFFRVYNAGKIVKYGILVRLLCESDTGYICNMEIYCGEGKRLLETIQTWSPYTDS WYHIYMDNYYNSVANCEALMKNKFRICGTIRKNRGIPKDFQTISLKKGETKFIRKNDILLQ VWQSKKPVYLISSIHSAEMEESQNIDRTSKKKIVKPNALIDYNKHMKGVDRADQYLSYYS ILRRTVKWTKRLAMYMINCALFNSYAVYKSVRQRKMGFKMFLKQTAIHWLTDDIPEDM DIVPDLQPVPSTSGMRAKPPTSDPPCRLSMDMRKHTLQAIVGSGKKKNILRRCRVCSVHK LRSETRYMCKFCNIPLHKGACFEKYHTLKNY (SEQ ID NO: 66) In embodiments, an enzyme capable of performing targeted genomic integration can act on an MLT left terminal end, or a sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto, where the nucleotide sequence of the MLT left terminal end (5′ to 3′) is as follows: ttaacacttggattgcgggaaacgagttaagtcggctcgcgtgaattgcgcgtactccgcgggagccgtcttaactcggttcatatagatttgcg gtggagtgcgggaaacgtgtaaactcgggccgattgtaactgcgtattaccaaatatttgtt (SEQ ID NO: 67) In embodiments, an enzyme capable of performing targeted genomic integration can act on an MLT right terminal end, or a sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto, wherein the nucleotide sequence of the MLT right terminal end (5′ to 3′) is as follows: aattatttatgtactgaatagataaaaaaatgtctgtgattgaataaattttcattttttacacaagaaaccgaaaatttcatttcaatcgaacccatactt caaaagatataggcattttaaactaactctgattttgcgcgggaaacctaaataattgcccgcgccatcttatattttggcgggaaattcacccgac accgtagtgttaa (SEQ ID NO: 68) Other forms of enzyme capable of performing targeted genomic integration include the following mobile element enzymes / transposases. Trichnoplusia ni MGSSLDDEHILSALLQSDDELVGEDSDSEISDHVSEDDVQSDTEEAFIDEVHEVQPTSSGSE ILDEQNVIEQPGSSLASNKILTLPQRTIRGKNKHCWSTSKSTRRSRVSALNIVRSQRGPTRM CRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESMTGATFRDTNEDEIYAFFGILVMTA VRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKSIRPTLRENDVFTPVRKI WDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRMYIPNKPSKYGIKILMMCDSGTKYMIN GMPYLGRGTQTNGVPLGEYYVKELSKPVRGSCRNITCDNWFTSIPLAKNLLQEPYKLTIV GTVRSNKREIPEVLKNSRSRPVGTSMFCFDGPLTLVSYKPKPAKMVYLLSSCDEDASINES TGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRKTNRWPMALLYGMINIACINSFIIYSHN VSSKGEKVQSRKKFMRNLYMSLTSSFMRKRLEAPTLKRYLRDNISNILPNEVPGTSDDSTE EPVTKKRTYCTYCPSKIRRKANASCKKCKKVICREHNIDMCQSCF (SEQ ID NO: 69) Pteropus vampyrus MSNPRKRSIPTCDVNFVLEQLLAEDSFDESDFSEIDDSDDFSDSASEDYTVRPPSDSESDGN SPTSADSGRALKWSTRVMIPRQRYDFTGTPGRKVDVSDTTDPLQYFELFFTEELVSKITSE MNAQAALLASKPPGPKGFSRMDKWKDTDNDELKVFFAVMLLQGIVQKPELEMFWSTRP LLDIPYLRQIMTGERFLLLLRCLHFVNNSSISAGQSKAQISLQKIKPVFDFLVNKFSTVYTPN RNIAVDESLMLFKGRLAMKQYIPTKMNLKDSADGLK (SEQ ID NO: 70) Myotis myotis (“2a”) MDLRCQHTVLSIRESRGLLPNLKMKTSRMKKGDIIFSRKGDILLLAWKDKRVVRMISIHDT SVSTTGKKNRKTGENIVKPACIKEYNAHMKGVDRADQFLSCCSILRKMMKWTKKVVLY LINCGLFNSFRVYNVLNPQAKMKYKQFLLSVARDWIMDDNNEGSPEPETNLSSPSPGGAR RAPRKDPPKRLSGDMKQHEPTCIPASGKKKFPTRACRVCAHGKRSESRYLCKFCLVPLHR GKCFTQYHTLKKY (SEQ ID NO: 71) Myotis myotis (“1”) MKAFLGVILNMGVLNHPNLQSYWSMDFESHIPFFRSVFKRERFLQIFWMLHLKNDQKSSK DLRTRTEKVNCFLSYLEMKFRERFCPGREIAVDEAVVGFKGKIHFITYNPKKPTKWGIRLY VLSDSKCGYVHSFVPYYGGITSETLVRPDLPFTSRIVLELHERLKNSVPGSQGYHFFTDRY YTSVTLAKELFKEKTHLTGTIMPNRKDNPPVIKHQKLKKGEIVAFRDENVMLLAWKDKRI VTLSTWDSETESVERRVGGGKEIVLKPKVVTNYTKFMGGVDIADYTSTYCFMRKTLKW WRTLFFWGLEVSVVNSYILYKECQKRKNEKPITHVKFIRKLVHDLVGEFRDGTLTSRGRL LSTNLEQRLDGKLHIITPHPNKKHKDCVVCSNRKIKGGRRETIYICETCECKPGLHVGECF KKYHTMKNYRD (SEQ ID NO: 72) Myotis lucifugus (“2”) MPSLRKRKETNETDTLPEVFNDNLSDIPSEIEDADDCFDDSGDDSTDSTDSEIIRPVRKRKV AVLSSDSDTDEATDNCWSEIDTPPRLQMFEGHAGVTTFPSQCDSVPSVTNLFFGDELFEML CKELSNYHDQTAMKRKTPSRTLKWSPVTQKDIKKFLGLIILMGQTRKDSLKDYWSTDPLI CTPIFPQTMSRHRFEQIWTFWHFNDNAKMDSRSGRLFKIQPVLDYFLHKFRTIYKPKQQLS LDEGMIPWRGRFKFRTYNPAKITKYGLLVRMVCESDTGYICSMEIYTAEGRKLQETVLSV LGPYLGIWHHIYQDNYYNATSTAELLLQNKTRVCGTIRESRGLPPNLEMKTSRMKKGDIIF SRKGDILLLAWKDKRVVRMISTIHDTSVSTTGKKNRKTGENIVKPTCIKEYNAHMKGVDR ADQFLSCCSILRKTMKWTKKVVLYLINCGLFNSFRVYNVLNPQAKMKYKQFLLSVARDW ITDDNNEGSPEPETNLSSPSPGGARRAPRKDPPKRLSGDMKQHEPTCIPASGKKKFPTRAC RVCAAHGKRSESRYLCKFCLVPLHRGKCFTQYHTLKKYMDLRCQHTVLSTVGRGYSVLA RFKPRTNERTGSSHCHVQVPAGGQGPPSTIIANGCGCKLEPMVRTRSPTCLVIEFGCM (SEQ ID NO: 73) Myotis myotis (“2”) MPSLRKRKETNETDTLPEVFNDNLSDIPSEIEDADDCFDDSGDDSTDSTESEIIRPVRKRKV AVLSSDSNTDEATDNCWSEIDTPPRLQMFEGHAGVTTFPSQCDSVPSVTNLFFGDELFEML CKELSNYHDQTAMKRKTPSRTLKWSPVTQKDIKKFLGLIILMGQTRKDSWKDYWSTDPLI CTPIFPQTMSRHRFEQIWTFWHFNDNAKMDSCSGRLFKIQPVLDYFLHKFRTIYKPKQQLS LDEGMIPWRGRLKFTYNPAITKYGLLVRMVCESDTGYICNMEIYTAERKKLQETVLSVLG PYLGIWHHIYQDNYYNATSTAELLLQNKTRVCGTIRESRGLPPNLKMKTSRMKKGDIIFSR KGDILLLAWKDKRVVRMISTIHDTSVSTTGKKNRKTGENIVKPTCIKEYNAHMKGVDRA DQFLSCCSILRKTTKWTKKVVLYLINCGLFNSFRVYNILNPQAKMKYKQFLLSVARDWIT DDNNEGSPEPETNLSSPSSGGARRAPRKDQPKRLSGDMKQHEPTCIPASGKKKFPTACRV CAAHGKRSESRYLRKFCFVPLRGKCFMYHTLKKYSELFSLIVVSKIQNVIIYKTTKVYMRY VMRSHCPLSFLVFAPSVKDRSRVFSFFTRHLLWTLDVNTLSCPHRMKRSHWWKPCRSIYE KLYNCTNP (SEQ ID NO: 74) Myotis myotis (“2b”) MDLRCQHTVLSIRESRGLPPNLKMKTSRMKKGDIIFSRKGDILLLAWKDKRVVRMISTIHD TSVSTTGKKNRKTGENIVKPACIKEYNAHMKGVDRADQFLSCCSILRKTMKWTKKVVLY LINCGLFNSFRVYNVLNPQAKMKYKQFLLSVARDWITDDNNEGSPEPETNLSSPSPGGAR RAPRKDPPKRLSGDMKQHEPTCIPASGKKKFPTRACRVCAAHGKRSESRYLCKFCLVPLH RGKCFTQYHTLKKY (SEQ ID NO: 75) In embodiments, an enzyme capable of performing targeted genomic integration is any type of an enzyme that causes a donor DNA (e.g., a CAR-encoding and shRNA-encoding nucleic acid molecule as disclosed herein) to be inserted from one location (e.g., without limitation, donor DNA) to a specific site and / or locus in a cell and / or subject's genome. In embodiments, the enzyme capable of performing targeted genomic integration is a recombinase. In embodiments, the recombinase is an integrase. In embodiments, the enzyme is a mobile element enzyme. In embodiments, the recombinase is an integrase or a mobile element enzyme. In embodiments, the mobile element enzyme is an engineered mammalian mobile element enzyme. In embodiments, the mobile element enzyme is a mammal-derived, helper RNA mobile element enzyme. Messenger RNA (mRNA) is an effective alternative to DNA as a source of a mobile element enzyme for targeting somatic cells and tissues, given that RNA is a safer alternative to DNA as a source of a mobile element enzyme for somatic gene therapy applications. See, e.g., Wilber et al., Mol. Ther. 13, 625-630 (2006). Successful use of in vitro-transcribed mRNA as a transient source of mobile element enzyme and subsequent transposition in cultured human cells and in live mice was previously reported for Sleeping Beauty mobile element enzyme. See id. It was demonstrated that in vitro-transcribed, UTR-stabilized mobile element enzyme-encoding mRNA can be used as a source of mobile element enzyme for Sleeping Beauty-mediated transposition in cultured somatic cells. Id. Also, Hoerr et al. reported that a specific cytotoxic T cell response and circulating antigen- specific antibodies were detected after administration of in vitro-transcribed, UTR-stabilized, and protamine-condensed bacterial lacZ mRNA into the ear pinna of Balb / C mice. Hoerr et al., Eur. J. lmmunol.2000; 30: 1-7; see also Wilber et al. (2006). In embodiments, the mobile element enzyme is a mammal-derived, DNA mobile element enzyme. In embodiments, the mobile element enzyme is a chimeric mobile element enzyme. In embodiments, the enzyme capable of performing targeted genomic integration is a mobile element enzyme, and the mobile element enzyme comprises (a) a targeting element which is or comprises a gene-editing system, and (b) a mobile element enzyme that is capable of inserting the donor DNA comprising a transgene, a chimeric CAR, and / or one or more shRNA-encoding sequence(s) optionally at a TA dinucleotide site or a TTAA tetranucleotide site, optionally in a genomic safe harbor site (GSHS), as known in the art. In some embodiments, a nucleic acid molecule / cassette as disclosed herein is inserted into a GSHS location in a host genome. GSHSs can be defined as loci well-suited for gene transfer, as integrations within these sites are not associated with adverse effects such as proto-oncogene activation, tumor suppressor inactivation, or insertional mutagenesis. GSHSs can be defined by the following criteria: 1) distance of at least 50 kb from the 5′ end of any gene, (2) distance of at least 300 kb from any cancer-related gene, (3) distance of at least 300 kb from any microRNA (miRNA), (4) location outside a transcription unit, and (5) location outside ultra-conserved regions (UCRs) of the human genome. See Papapetrou et al. Nat Biotechnol 2011; 29:73-8; Bejerano et al. Science 2004; 304:1321-5. In embodiments, the enzyme is derived from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Myotis / ucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, Molossus molossus, Pan troglodytes, or Homo sapiens. In embodiments, the enzyme is an engineered version, including but not limited to hyperactive forms, of an enzyme derived from Bombyx mori, Xenopus tropica / is, Trichoplusia ni, Myotis / ucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, Molossus molossus, Pan troglodytes, or Homo sapiens. In embodiments, the enzyme is a mobile element enzyme derived from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Myotis / ucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, Molossus molossus, Pan troglodytes, or Homo sapiens. In embodiments, the enzyme is an engineered version, including but not limited to hyperactive forms, of a mobile element enzyme derived from Bombyx mori, Xenopus tropica / is, Trichoplusia ni, Myotis lucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, Molossus molossus, Pan troglodytes, or Homo sapiens. In embodiments, the mobile element enzyme is from one or more of the Tn1, Tn2, Tn3, Tn5, Tn7, Tn9, Tn10, Tn552, Tn903, Tn1000 / Gamma-delta, Tn / O, tnsA, tnsB, tnsC, tniQ, IS10, ISS, IS911, Minos, Sleeping beauty, piggyBac, Tol2, Mos1, Himar1, Hermes, Tol2, Minos, Tel, P- element, MuA, Ty1, Chapaev, transib, Tc1 / mariner, or Tc3 donor DNA system, or biologically active fragments variants thereof, inclusive of hyperactive mutants. In some embodiments, the transposase is from a Sleeping Beauty transposon system (see, e.g., Cell.1997; 91:501-510), e.g. a hyperactive form of Sleeping Beauty (hypSB), e.g. SB100X (see Gene Therapy volume 18, pages 849-856(2011), or a piggyBac (PB) transposon system (see, e.g. Trends Biotechnol. 2015 September; 33(9):525-33, which is incorporated herein by reference in its entirety), e.g. a hyperactive form of PB transposase (hypPB), e.g. with seven amino acid substitutions (e.g. I30V, S103P, G165S, M282V, S509G, N570S, N538K on mPB, or functional equivalents in non-mPB, see Mol Ther Nucleic Acids.2012 October; 1(10): e50, which is incorporated herein by reference in its entirety); see also Yusa et al., PNAS Jan. 25, 2011108 (4) 1531-1536; Voigt et al., Nature Communications volume 7, Article number: 11126 (2016). The piggyBac transposases belong to the IS4 transposase family. De Palmenaer et al., BMC Evolutionary Biology.2008; 8:18. doi: 10.1186 / 1471-2148-8-18. The piggyBac family includes a large diversity of transposons, and any of these transposons can be used in embodiments of the present disclosure. See, e.g., Bouallègue et al., Genome Biol Evol.2017; 9(2):323-339. The founding member of the piggyBac (super)family, insect piggyBac, was originally identified in the cabbage looper moth (Trichoplusiani ni) and studied both in vivo and in vitro. Insect piggyBac is known to transpose by a canonical cut-and-paste mechanism promoted by an element-encoded transposase with a catalytic site resembling the RNase H fold shared by many recombinases. The insect piggyBac transposon system has been shown to be highly active in a wide range of animals, including Drosophila and mice, where it has been developed as a powerful tool for gene tagging and genome engineering. Other transposons affiliated to the piggyBac superfamily are common in arthropods and vertebrates including Xenopus and Bombyx. Mammalian piggyBac transposons and transposases, including hyperactive mammalian piggyBac variants, which can be used in embodiments of the present disclosure, are described, e.g., in International Application WO2010085699, which is incorporated herein by reference in its entirety. In some embodiments, the transposase is from a MLT transposon system that is based on a cut-and-paste MLT element obtained from the little brown bat (Myotis lucifugus) or other bat transposases, such as Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pipistrellus kuhlii and Molossus molossus. See Mitra et al., Proc Natl Acad Sci USA.2013 Jan. 2; 110(1):234-9; Jebb et al., Nature, volume 583, pages 578-584 (2020), which is incorporated by reference herein in its entirety. In some embodiments, hyperactive forms of a bat transposase is used. The MLT transposase has been shown to be capable of transposition in bat, human, and yeast cells. The hyperactive forms of the MLT transposase enhance the transposition process. In addition, chimeric MLT transposases are capable of site-specific excision without genomic integration. Furthermore, in embodiments, the engineered and / or corrected MLT transposase is used that has certain mutations relative to the wild-type MLT transposase. In embodiments, hyperactive forms of the corrected MLT transposase are used. In embodiments, as also detailed further above, the transposase enzyme is derived from any of Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Myotis lucifugus, Pipistrellus kuhlii, Pteropus vampyrus, and Molossus molossus. In embodiments, the transposase enzyme is derived from any of Trichoplusia ni, Myotis lucifugus, Myotis myotis, or Pteropus vampyrus. Lipid Nanoparticles (LNPs) In certain embodiments, contemporaneous expression of (1) an enzyme capable of performing targeted genomic integration and (2) a CAR-encoding and shRNA-encoding nucleic acid molecule of the instant disclosure that is flanked by recognition sequences (e.g., terminal repeats) for the enzyme capable of performing targeted genomic integration are achieved in a target cell (e.g., an immune cell, e.g., a T cell) via lipid nanoparticle (LNP)-mediated administration of both a nucleic acid encoding for the enzyme capable of performing targeted genomic integration and the CAR- encoding and shRNA-encoding nucleic acid molecule. Any of a variety of LNPs known in the art, or for that matter non-LNP-mediated modes of transfection and / or contact / administration of peptides and / or nucleic acid molecules for expression and achievement of genomic integration, can be employed to make the engineered cells of the instant disclosure. In embodiments, the nucleic acid encoding the enzyme and the the CAR-encoding and shRNA-encoding nucleic acid molecule are in the form of a co-formulation incorporated into or associated with the same LNP. In embodiments, the LNP is selected from 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), a cationic cholesterol derivative mixed with dimethylaminoethane-carbamoyl (DC-Chol), phosphatidylcholine (PC), triolein (glyceryl trioleate), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [carboxy(polyethylene glycol)-2000] (DSPE-PEG), 1,2-dimyristoyl-rac-glycero-3- methoxypolyethyleneglycol-2000 (DMG-PEG 2K), and 1,2 distearol-sn-glycerol- 3phosphocholine (DSPC) and / or comprising of one or more molecules selected from polyethylenimine (PEI) and poly(lactic-co-glycolic acid) (PLGA), and N-Acetylgalactosamine (GalNAc). In embodiments, an LNP is as described, e.g., in Patel et al., J Control Release 2019; 303:91-100. The LNP can comprise one or more of a structural lipid (e.g., DSPC), a PEG-conjugated lipid (COM-PEG), a cationic lipid (MC3), cholesterol, and a targeting ligand (e.g., GalNAc). In embodiments, a nanoparticle is a particle having a diameter of less than about 1000 nm. In embodiments, nanoparticles of the present disclosure have a greatest dimension (e.g., diameter) of about 500 nm or less, or about 400 nm or less, or about 300 nm or less, or about 200 nm or less, or about 100 nm or less. In embodiments, nanoparticles of the present invention have a greatest dimension ranging between about 50 nm and about 150 nm, or between about 70 nm and about 130 nm, or between about 80 nm and about 120 nm, or between about 90 nm and about 110 nm. In embodiments, the nanoparticles of the present disclosure have a greatest dimension (e.g., a diameter) of about 100 nm. In some aspects, the cell in accordance with the present disclosure is prepared via an in vivo genetic modification method. In embodiments, a genetic modification in accordance with the present disclosure is performed via an ex vivo method. In some aspects, the cell in accordance with the present disclosure is prepared by contacting a cell with an enzyme capable of performing targeted genomic integration (e.g., without limitation, a mammalian mobile element enzyme) in vivo. In embodiments, the cell is contacted with the enzyme ex vivo. In embodiments, the mobile element enzyme(s) used herein feature reduced insertional mutagenesis or oncogenesis as compared to non- chimeric mobile element enzyme. Immune Effector Cells As contemplated herein, the CARs of the present disclosure are expressed by an immune effector cell, for example a T-cell, and administered to a subject in order to treat a disease or disorder, for example, a cancer. Among the cell types that may be used to express the CARs of the present disclosure include, but are not limited to, T-cells, NK cells, CD4+ T- cells, CD8+ cells, and stem cells, such as an induced pluripotent stem cell (iPS cell). In one embodiment, the cell is an autologous T-cell. In one embodiment, the cell shows anti-tumor activity when cross-reacted with a tumor cell containing an antigen capable of being bound by the extracellular ligand binding domain. The generation of CAR T-cells is known in the art. For example, see Wang et al, “Clinical manufacturing of CAR T cells: foundation of a promising therapy,” Oncolytics (2016) 3:1-7 (and incorporated herein). In general, the CAR T-cells of the disclosure can be generated by introducing a nucleic acid molecule / construct as disclosed herein, optionally within a lentiviral vector and / or with an integrating enzyme, the construct and / or lentiviral vector including a desired CAR and / or CAR system, for example a CAR comprising anti-CD19, CD8α hinge and transmembrane domain, human CD28 and CD3zeta signaling domains, into the cells. The CAR T-cells of the disclosure are able to replicate in vivo resulting in long-term persistence that can lead to sustained tumor control. The modified cell (e.g., T cells) described herein may be included in a composition for immunotherapy. The composition may include a pharmaceutical composition and further include a pharmaceutically acceptable carrier. A therapeutically effective amount of the pharmaceutical composition comprising the modified T cells may be administered. In one aspect, the disclosure includes a method for adoptive cell transfer therapy comprising administering to a subject in need thereof a population of modified cells of the present disclosure, wherein the cells are immune cells or precursor cells thereof (e.g., T cells). Methods for administration of immune cells for adoptive cell therapy are known and may be used in connection with the provided methods and compositions. For example, adoptive immune cell therapy methods are described, e.g., in US Patent Application Publication No.2003 / 0170238 to Gruenberg et al; U.S. Pat. No. 4,690,915 to Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85). See, e.g., Themeli et al. (2013) Nat Biotechnol. 31(10): 928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1): 84-9; Davila et al. (2013) PLoS ONE 8(4): e61338; Lee et al., Int J. Mol Sci. (2021) 22(9):4590; Banerjee et al., JCO Clin Cancer Inform. (2021) 5:668- 678; Robbins et al., Stem Cell Res Ther. (2021) 12(1):350; Wrona et al., Int J Mol Sci. (2021) 22(11):5899; Atrash and Moyo, Onco Targets Ther. (2021) 14:2185-2201; Martinez Bedoya et al., Front Immunol. (2021) 12:640082; Morgan et al., Front Immunol. (2020) 11:1965; Chicaybam et al., Cancers (Basel) (2020) 12(9):2360; and Rafiq et al., Nat Rev Clin Oncol. (2020) 17(3):147-167. In some embodiments, the cell therapy, e.g., adoptive T cell therapy is carried out by autologous transfer, in which the cells are isolated and / or otherwise prepared from the subject who is to receive the cell therapy, or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., patient, in need of a treatment and the cells, following isolation and processing are administered to the same subject. In some embodiments, the cell therapy, e.g., adoptive T cell therapy, is carried out by allogeneic transfer, in which the cells are isolated and / or otherwise prepared from a subject other than a subject who is to receive or who ultimately receives the cell therapy, e.g., a first subject. In such embodiments, the cells then are administered to a different subject, e.g., a second subject, of the same species. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject. In some embodiments, the subject has been treated with a therapeutic agent targeting the disease or condition, e.g., the tumor, prior to administration of the cells or composition containing the cells. In some aspects, the subject is refractory or non-responsive to the other therapeutic agent. In some embodiments, the subject has persistent or relapsed disease, e.g., following treatment with another therapeutic intervention, including chemotherapy, radiation, and / or hematopoietic stem cell transplantation (HSCT), e.g., allogenic HSCT. In some embodiments, the administration effectively treats the subject despite the subject having become resistant to another therapy. In some embodiments, the subject is responsive to the other therapeutic agent, and treatment with the therapeutic agent reduces disease burden. In some aspects, the subject is initially responsive to the therapeutic agent, but exhibits a relapse of the disease or condition over time. In some embodiments, the subject has not relapsed. In some such embodiments, the subject is determined to be at risk for relapse, such as at a high risk of relapse, and thus the cells are administered prophylactically, e.g., to reduce the likelihood of or prevent relapse. In some aspects, the subject has not received prior treatment with another therapeutic agent. In some embodiments, the subject has persistent or relapsed disease, e.g., following treatment with another therapeutic intervention, including chemotherapy, radiation, and / or hematopoietic stem cell transplantation (HSCT), e.g., allogenic HSCT. In some embodiments, the administration effectively treats the subject despite the subject having become resistant to another therapy. The modified immune cell of the present disclosure can be administered to an animal, preferably a mammal, even more preferably a human, to treat a cancer. In addition, the cells of the present disclosure can be used for the treatment of any condition related to a cancer, especially a cell-mediated immune response against a tumor cell(s), where it is desirable to treat or alleviate the disease. The types of cancers to be treated with the modified cells or pharmaceutical compositions of the disclosure include certain leukemia or lymphoid malignancies, benign and malignant tumors, and malignancies e.g., sarcomas, carcinomas, and melanomas. Exemplary cancers include but are not limited to B-cell malignancies (such as B-cell lymphomas and leukemias and the like), lung cancer, non-small cell lung cancer, small cell lung cancer, Merkel cell carcinoma, melanoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, urothelial carcinoma, gastric cancer, cervical cancer, cutaneous squamous cell carcinoma, renal cell carcinoma, breast cancer, triple-negative breast cancer, colon cancer, esophagus cancer, stomach cancer, liver cancer, kidney cancer, pancreatic cancer, prostate cancer, brain cancer, lung adenocarcinoma, glioblastoma, hepatocellular carcinoma, gallbladder cancer, cervical cancer, cervical squamous cell carcinoma, colorectal cancer, ovarian cancer, and renal cancer. The cancers may be non-solid tumors (such as hematological tumors) or solid tumors. Adult tumors / cancers and pediatric tumors / cancers are also included. In one embodiment, the cancer is a solid tumor or a hematological tumor. The cells to be administered may be autologous, with respect to the subject undergoing therapy. The administration of the cells of the disclosure may be carried out in any convenient manner known to those of skill in the art. The cells of the present disclosure may be administered to a subject by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. The compositions described herein may be administered to a patient transarterially, subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous (i.v.) injection, or intraperitoneally. In other instances, the cells of the disclosure are injected directly into a site of inflammation in the subject, a local disease site in the subject, a lymph node, an organ, a tumor, and the like. In some embodiments, the cells are administered at a desired dosage, which in some aspects includes a desired dose or number of cells or cell type(s) and / or a desired ratio of cell types. Thus, the dosage of cells in some embodiments is based on a total number of cells (or number per kg body weight) and a desired ratio of the individual populations or sub-types, such as the CD4+to CD8+ratio. In some embodiments, the dosage of cells is based on a desired total number (or number per kg of body weight) of cells in the individual populations or of individual cell types. In some embodiments, the dosage is based on a combination of such features, such as a desired number of total cells, desired ratio, and desired total number of cells in the individual populations. In some embodiments, the populations or sub-types of cells, such as CD8+and CD4+T cells, are administered at or within a tolerated difference of a desired dose of total cells, such as a desired dose of T cells. In some aspects, the desired dose is a desired number of cells or a desired number of cells per unit of body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is at or above a minimum number of cells or minimum number of cells per unit of body weight. In some aspects, among the total cells, administered at the desired dose, the individual populations or sub-types are present at or near a desired output ratio (such as CD4+to CD8+ratio), e.g., within a certain tolerated difference or error of such a ratio. In some embodiments, the cells are administered at or within a tolerated difference of a desired dose of one or more of the individual populations or sub-types of cells, such as a desired dose of CD4+cells and / or a desired dose of CD8+cells. In some aspects, the desired dose is a desired number of cells of the sub-type or population, or a desired number of such cells per unit of body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is at or above a minimum number of cells of the population or subtype, or minimum number of cells of the population or sub-type per unit of body weight. Thus, in some embodiments, the dosage is based on a desired fixed dose of total cells and a desired ratio, and / or based on a desired fixed dose of one or more, e.g., each, of the individual sub-types or sub-populations. Thus, in some embodiments, the dosage is based on a desired fixed or minimum dose of T cells and a desired ratio of CD4+to CD8+cells, and / or is based on a desired fixed or minimum dose of CD4+and / or CD8+cells. Types of cancers to be treated with the CAR-expressing cells of the disclosure include, but are not limited to, carcinoma, blastoma, and sarcoma, and certain leukemia or lymphoid malignancies, benign and malignant tumors, and malignancies e.g., sarcomas, carcinomas, and melanomas. Adult tumors / cancers and pediatric tumors / cancers are also included. Hematologic cancers are cancers of the blood or bone marrow. Examples of hematological (or hematogenous) cancers include leukemias, including acute leukemias (such as acute lymphocytic leukemia, acute myelocytic leukemia, acute myelogenous leukemia and myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia), chronic leukemias (such as chronic myelocytic (granulocytic) leukemia, chronic myelogenous leukemia, and chronic lymphocytic leukemia), polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma (indolent and high grade forms), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia and myelodysplasia. Other hematological cancers include T-cell or NK-cell lymphoma, for example, but not limited to: peripheral T-cell lymphoma; anaplastic large cell lymphoma, for example anaplastic lymphoma kinase (ALK) positive, ALK negative anaplastic large cell lymphoma, or primary cutaneous anaplastic large cell lymphoma; angioimmunoblastic lymphoma; cutaneous T-cell lymphoma, for example mycosis fungoides, Sézary syndrome, primary cutaneous anaplastic large cell lymphoma, primary cutaneous CD30+ T-cell lymphoproliferative disorder; primary cutaneous aggressive epidermotropic CD8+ cytotoxic T-cell lymphoma; primary cutaneous gamma-delta T- cell lymphoma; primary cutaneous small / medium CD4+ T-cell lymphoma, and lymphomatoid papulosis; Adult T-cell Leukemia / Lymphoma (ATLL); Blastic NK-cell Lymphoma; Enteropathy- type T-cell lymphoma; Hematosplenic gamma-delta T-cell Lymphoma; Lymphoblastic Lymphoma; Nasal NK / T-cell Lymphomas; Treatment-related T-cell lymphomas; for example lymphomas that appear after solid organ or bone marrow transplantation; T-cell prolymphocytic leukemia; T-cell large granular lymphocytic leukemia; Chronic lymphoproliferative disorder of NK- cells; Aggressive NK cell leukemia; Systemic EBV+ T-cell lymphoproliferative disease of childhood (associated with chronic active EBV infection); Hydroa vacciniforme-like lymphoma; Adult T-cell leukemia / lymphoma; Enteropathy-associated T-cell lymphoma; Hepatosplenic T-cell lymphoma; or Subcutaneous panniculitis-like T-cell lymphoma. In one embodiment, the CAR expressing cells can be used in an effective amount to treat a host, for example a human, with a lymphoma or lymphocytic or myelocytic proliferation disorder or abnormality. For example, the CAR expressing cells as described herein can be administered to a host suffering from a Hodgkin Lymphoma or a Non-Hodgkin Lymphoma. For example, the host can be suffering from a Non-Hodgkin Lymphoma such as, but not limited to: an AIDS-Related Lymphoma; Anaplastic Large-Cell Lymphoma; Angioimmunoblastic Lymphoma; Blastic NK-Cell Lymphoma; Burkitt’s Lymphoma; Burkitt-like Lymphoma (Small Non-Cleaved Cell Lymphoma); Chronic Lymphocytic Leukemia / Small Lymphocytic Lymphoma; Cutaneous T-Cell Lymphoma; Diffuse Large B- Cell Lymphoma; Enteropathy-Type T-Cell Lymphoma; Follicular Lymphoma; Hepatosplenic Gamma-Delta T-Cell Lymphoma; Lymphoblastic Lymphoma; Mantle Cell Lymphoma; Marginal Zone Lymphoma; Nasal T-Cell Lymphoma; Pediatric Lymphoma; Peripheral T- Cell Lymphomas; Primary Central Nervous System Lymphoma; T-Cell Leukemias; Transformed Lymphomas; Treatment-Related T-Cell Lymphomas; or Waldenstrom's Macroglobulinemia. Alternatively, CAR expressing cells disclosed herein can be used in an effective amount to treat a host, for example a human, with a Hodgkin Lymphoma, such as, but not limited to: Nodular Sclerosis Classical Hodgkin’s Lymphoma (CHL); Mixed Cellularity CHL; Lymphocyte-depletion CHL; Lymphocyte-rich CHL; Lymphocyte Predominant Hodgkin Lymphoma; or Nodular Lymphocyte Predominant HL. Alternatively, CAR expressing cells disclosed herein can be used in an effective amount to treat a host, for example a human with a specific B-cell lymphoma or proliferative disorder such as, but not limited to: multiple myeloma; Diffuse large B cell lymphoma; Follicular lymphoma; Mucosa-Associated Lymphatic Tissue lymphoma (MALT); Small cell lymphocytic lymphoma; Mediastinal large B cell lymphoma; Nodal marginal zone B cell lymphoma (NMZL); Splenic marginal zone lymphoma (SMZL); Intravascular large B-cell lymphoma; Primary effusion lymphoma; or Lymphomatoid granulomatosis;; B-cell prolymphocytic leukemia; Hairy cell leukemia; Splenic lymphoma / leukemia, unclassifiable; Splenic diffuse red pulp small B-cell lymphoma; Hairy cell leukemia-variant; Lymphoplasmacytic lymphoma; Heavy chain diseases, for example, Alpha heavy chain disease, Gamma heavy chain disease, Mu heavy chain disease; Plasma cell myeloma; Solitary plasmacytoma of bone; Extraosseous plasmacytoma; Primary cutaneous follicle center lymphoma; T-cell / histiocyte rich large B-cell lymphoma; DLBCL associated with chronic inflammation; Epstein-Barr virus (EBV)+ DLBCL of the elderly; Primary mediastinal (thymic) large B-cell lymphoma; Primary cutaneous DLBCL, leg type; ALK+ large B-cell lymphoma; Plasmablastic lymphoma; Large B-cell lymphoma arising in HHV8-associated multicentric; Castleman disease; B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma; or B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma. In one embodiment, CAR expressing cells disclosed herein can be used in an effective amount to treat a host, for example a human with leukemia. For example, the host may be suffering from an acute or chronic leukemia of a lymphocytic or myelogenous origin, such as, but not limited to: Acute lymphoblastic leukemia (ALL); Acute myelogenous leukemia (AML); Chronic lymphocytic leukemia (CLL); Chronic myelogenous leukemia (CML); juvenile myelomonocytic leukemia (JMML); hairy cell leukemia (HCL); acute promyelocytic leukemia (a subtype of AML); large granular lymphocytic leukemia; or Adult T-cell chronic leukemia. In one embodiment, the patient suffers from an acute myelogenous leukemia, for example an undifferentiated AML (M0); myeloblastic leukemia (M1; with / without minimal cell maturation); myeloblastic leukemia (M2; with cell maturation); promyelocytic leukemia (M3 or M3 variant [M3V]); myelomonocytic leukemia (M4 or M4 variant with eosinophilia [M4E]); monocytic leukemia (M5); erythroleukemia (M6); or megakaryoblastic leukemia (M7). In one embodiment, CAR expressing cells disclosed herein can be used in an effective amount to treat a host, for example a human with a solid tumor. Examples include, but are not limited to, but are not limited to: estrogen-receptor positive, HER2-negative advanced breast cancer, late- line metastatic breast cancer, liposarcoma, non-small cell lung cancer, liver cancer, ovarian cancer, glioblastoma, refractory solid tumors, retinoblastoma positive breast cancer as well as retinoblastoma positive endometrial, vaginal and ovarian cancers and lung and bronchial cancers, adenocarcinoma of the colon, adenocarcinoma of the rectum, central nervous system germ cell tumors, teratomas, estrogen receptor-negative breast cancer, estrogen receptor-positive breast cancer, familial testicular germ cell tumors, HER2-negative breast cancer, HER2-positive breast cancer, male breast cancer, ovarian immature teratomas, ovarian mature teratoma, ovarian monodermal and highly specialized teratomas, progesterone receptor-negative breast cancer, progesterone receptor-positive breast cancer, recurrent breast cancer, recurrent colon cancer, recurrent extragonadal germ cell tumors, recurrent extragonadal non-seminomatous germ cell tumor, recurrent extragonadal seminomas, recurrent malignant testicular germ cell tumors, recurrent melanomas, recurrent ovarian germ cell tumors, recurrent rectal cancer, stage III extragonadal non- seminomatous germ cell tumors, stage III extragonadal seminomas, stage III malignant testicular germ cell tumors, stage III ovarian germ cell tumors, stage IV breast cancers, stage IV colon cancers, stage IV extragonadal non-seminomatous germ cell tumors, stage IV extragonadal seminoma, stage IV melanomas, stage IV ovarian germ cell tumors, stage IV rectal cancers, testicular immature teratomas, testicular mature teratomas, estrogen-receptor positive, HER2-negative advanced breast cancer, late-line metastatic breast cancer, liposarcoma, non-small cell lung cancer, liver cancer, ovarian cancer, glioblastoma, refractory solid tumors, retinoblastoma positive breast cancer as well as retinoblastoma positive endometrial, vaginal and ovarian cancers and lung and bronchial cancers, metastatic colorectal cancer, metastatic melanoma, or cisplatin-refractory, unresectable germ cell tumors, carcinoma, sarcoma, including, but not limited to, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, fibrosarcoma, myxosarcoma, chondrosarcoma, osteosarcoma, chordoma, malignant fibrous histiocytoma, hemangiosarcoma, angiosarcoma, lymphangiosarcoma. Mesothelioma, leiomyosarcoma, rhabdomyosarcoma, squamous cell carcinoma; epidermoid carcinoma, malignant skin adnexal tumors, adenocarcinoma, hepatoma, hepatocellular carcinoma, renal cell carcinoma, hypernephroma, cholangiocarcinoma, transitional cell carcinoma, choriocarcinoma, seminoma, embryonal cell carcinoma, glioma anaplastic; glioblastoma multiforme,, neuroblastoma, medulloblastoma, malignant meningioma, malignant schwannoma, neurofibrosarcoma, parathyroid carcinoma, medullary carcinoma of thyroid, bronchial carcinoid, pheochromocytoma, IsleT-cell carcinoma, malignant carcinoid, malignant paraganglioma, melanoma, Merkel cell neoplasm, cystosarcoma phylloide, salivary cancers, thymic carcinomas, bladder cancer, and Wilms tumor, a blood disorder or a hematologic malignancy, including, but not limited to, myeloid disorder, lymphoid disorder, leukemia, lymphoma, myelodysplastic syndrome (MDS), myeloproliferative disease (MPD), masT-cell disorder, and myeloma (e.g., multiple myeloma). In another embodiment, a CAR expressing cell disclosed herein can be used in an effective amount to treat a host, for example a human with an autoimmune disorder. Examples include, but are not limited to: Acute disseminated encephalomyelitis (ADEM); Addison's disease; Agammaglobulinemia; Alopecia areata; Amyotrophic lateral sclerosis (Also Lou Gehrig's disease; Motor Neuron Disease); Ankylosing Spondylitis; Antiphospholipid syndrome; Antisynthetase syndrome; Atopic allergy; Atopic dermatitis; Autoimmune aplastic anemia; Autoimmune arthritis; Autoimmune cardiomyopathy; Autoimmune enteropathy; Autoimmune granulocytopenia; Autoimmune hemolytic anemia; Autoimmune hepatitis; Autoimmune hypoparathyroidism; Autoimmune inner ear disease; Autoimmune lymphoproliferative syndrome; Autoimmune myocarditis; Autoimmune pancreatitis; Autoimmune peripheral neuropathy; Autoimmune ovarian failure; Autoimmune polyendocrine syndrome; Autoimmune progesterone dermatitis; Autoimmune thrombocytopenic purpura; Autoimmune thyroid disorders; Autoimmune urticarial; Autoimmune uveitis; Autoimmune vasculitis; Balo disease / Balo concentric sclerosis; Behçet's disease; Berger's disease; Bickerstaff's encephalitis; Blau syndrome; Bullous pemphigoid; Cancer; Castleman's disease; Celiac disease; Chagas disease; Chronic inflammatory demyelinating polyneuropathy; Chronic inflammatory demyelinating polyneuropathy; Chronic obstructive pulmonary disease; Chronic recurrent multifocal osteomyelitis; Churg-Strauss syndrome; Cicatricial pemphigoid; Cogan syndrome; Cold agglutinin disease; Complement component 2 deficiency; Contact dermatitis; Cranial arteritis; CREST syndrome; Crohn's disease; Cushing's Syndrome; Cutaneous leukocytoclastic angiitis; Dego's disease; Dercum's disease; Dermatitis herpetiformis; Dermatomyositis; Diabetes mellitus type 1; Diffuse cutaneous systemic sclerosis; Discoid lupus erythematosus; Dressler's syndrome; Drug-induced lupus; Eczema; Endometriosis; Enthesitis- related arthritis; Eosinophilic fasciitis; Eosinophilic gastroenteritis; Eosinophilic pneumonia; Epidermolysis bullosa acquisita; Erythema nodosum; Erythroblastosis fetalis; Essential mixed cryoglobulinemia; Evan's syndrome; Extrinsic and intrinsic reactive airways disease (asthma); Fibrodysplasia ossificans progressive; Fibrosing alveolitis (or Idiopathic pulmonary fibrosis); Gastritis; Gastrointestinal pemphigoid; Glomerulonephritis; Goodpasture's syndrome; Graves' disease; Guillain-Barré syndrome (GBS); Hashimoto's encephalopathy; Hashimoto's thyroiditis; Hemolytic anemia; Henoch-Schonlein purpura; Herpes gestationis (Gestational Pemphigoid); Hidradenitis suppurativa; Hughes-Stovin syndrome; Hypogammaglobulinemia; Idiopathic inflammatory demyelinating diseases; Idiopathic pulmonary fibrosis; Idiopathic thrombocytopenic purpura; IgA nephropathy; Immune glomerulonephritis; Immune nephritis; Immune pneumonitis; Inclusion body myositis; inflammatory bowel disease; Interstitial cystitis; Juvenile idiopathic arthritis aka Juvenile rheumatoid arthritis; Kawasaki's disease; Lambert-Eaton myasthenic syndrome; Leukocytoclastic vasculitis; Lichen planus; Lichen sclerosus; Linear IgA disease (LAD); Lupoid hepatitis aka Autoimmune hepatitis; Lupus erythematosus; Majeed syndrome; microscopic polyangiitis; Miller-Fisher syndrome; mixed connective tissue disease; Morphea; Mucha-Habermann disease aka Pityriasis lichenoides et varioliformis acuta; Multiple sclerosis; Myasthenia gravis; Myositis; Ménière's disease; Narcolepsy; Neuromyelitis optica (also Devic's disease); Neuromyotonia; Occular cicatricial pemphigoid; Opsoclonus myoclonus syndrome; Ord's thyroiditis; Palindromic rheumatism; PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcus); Paraneoplastic cerebellar degeneration; Paroxysmal nocturnal hemoglobinuria (PNH); Parry Romberg syndrome; Pars planitis; Parsonage- Turner syndrome; Pemphigus vulgaris; Perivenous encephalomyelitis; Pernicious anaemia; POEMS syndrome; Polyarteritis nodosa; Polymyalgia rheumatic; Polymyositis; Primary biliary cirrhosis; Primary sclerosing cholangitis; Progressive inflammatory neuropathy; Psoriasis; Psoriatic arthritis; pure red cell aplasia; Pyoderma gangrenosum; Rasmussen's encephalitis; Raynaud phenomenon; Reiter's syndrome; relapsing polychondritis; restless leg syndrome; retroperitoneal fibrosis; rheumatic fever; rheumatoid arthritis; Sarcoidosis; Schizophrenia; Schmidt syndrome; Schnitzler syndrome; Scleritis; Scleroderma; Sclerosing cholangitis; serum sickness; Sjögren's syndrome; Spondyloarthropathy; Stiff person syndrome; Still's disease; Subacute bacterial endocarditis (SBE); Susac's syndrome; Sweet's syndrome; Sydenham chorea; sympathetic ophthalmia; systemic lupus erythematosus; Takayasu's arteritis; temporal arteritis (also known as "gianT-cell arteritis"); thrombocytopenia; Tolosa-Hunt syndrome; transverse myelitis; ulcerative colitis; undifferentiated connective tissue disease; undifferentiated spondyloarthropathy; urticarial vasculitis; vasculitis; vitiligo; viral diseases such as Epstein Barr Virus (EBV), Hepatitis B, Hepatitis C, HIV, HTLV 1, Varicella-Zoster Virus (VZV) and Human Papilloma Virus (HPV); or Wegener's granulomatosis. In some embodiments, the autoimmune disease is an allergic condition, including those from asthma, food allergies, atopic dermatitis, and rhinitis. Solid tumors are abnormal masses of tissue that usually do not contain cysts or liquid areas. Solid tumors can be benign or malignant. Different types of solid tumors are named for the type of cells that form them (such as sarcomas, carcinomas, and lymphomas). Examples of solid tumors, such as sarcomas and carcinomas, include fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, and other sarcomas, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, lymphoid malignancy, pancreatic cancer, breast cancer, lung cancers, ovarian cancer, prostate cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, pheochromocytomas sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, Wilms' tumor, cervical cancer, testicular tumor, seminoma, bladder carcinoma, melanoma, and CNS tumors (such as a glioma (such as brainstem glioma and mixed gliomas), glioblastoma (also known as glioblastoma multiforme) astrocytoma, CNS lymphoma, germinoma, medulloblastoma, Schwannoma craniopharyogioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, neuroblastoma, retinoblastoma and brain metastases). In one embodiment, the antigen binding moiety portion of the CAR of the disclosure is designed to treat a particular cancer. For example, a CAR designed to target CD19 can be used to treat cancers and disorders including but are not limited to pre-B ALL (pediatric indication), adult ALL, mantle cell lymphoma, diffuse large B-cell lymphoma, salvage post allogeneic bone marrow transplantation, and the like. In another embodiment, the CAR can be designed to target CD22 to treat diffuse large B- cell lymphoma. In one embodiment, cancers and disorders include but are not limited to pre-B ALL (pediatric indication), adult ALL, mantle cell lymphoma, diffuse large B-cell lymphoma, salvage post allogeneic bone marrow transplantation, and the like can be treated using a combination of CARs that target CD19, CD20, CD22, and ROR1. In one embodiment, the CAR can be designed to target mesothelin to treat mesothelioma, pancreatic cancer, ovarian cancer, and the like. In one embodiment, the CAR can be designed to target CD33 / IL3Ra to treat acute myelogenous leukemia and the like. In one embodiment, the CAR can be designed to target CD30 to treat lymphoma, for example Hodgkin lymphoma, and the like. In one embodiment, the CAR can be designed to target c-Met to treat triple negative breast cancer, non-small cell lung cancer, and the like. In one embodiment, the CAR can be designed to target PSMA to treat prostate cancer and the like. In one embodiment, the CAR can be designed to target Glycolipid F77 to treat prostate cancer and the like. In one embodiment, the CAR can be designed to target EGFRvIII to treat gliobastoma and the like. In one embodiment, the CAR can be designed to target GD-2 to treat neuroblastoma, melanoma, and the like. In one embodiment, the CAR can be designed to target NY-ESO-1 TCR to treat myeloma, sarcoma, melanoma, and the like. In one embodiment, the CAR can be designed to target MAGE A3 TCR to treat myeloma, sarcoma, melanoma, and the like. In one embodiment, the CAR can be designed to target CEA to treat colorectal cancer and the like. In one embodiment, the CAR can be designed to target erb-B2, erb-B3, and / or erb-B4 to treat breast cancer, and the like. In one embodiment, the CAR can be designed to target IL-13R-a2 to treat glioma, glioblastoma, or medulloblastoma, and the like. However, the disclosure should not be construed to be limited to solely to the antigen targets and diseases disclosed herein. Rather, the disclosure should be construed to include any antigenic or ligand target that is associated with a disease where a CAR can be used to treat the disease or disorder. The CAR-expressing cells of the disclosure may also serve as a type of vaccine for ex vivo immunization and / or in vivo therapy in a mammal. Optionally, the mammal is a human. With respect to ex vivo immunization, at least one of the following occurs in vitro prior to administering the cell into a mammal: i) expansion of the cells, ii) introducing a nucleic acid encoding a CAR to the cells, and / or iii) cryopreservation of the cells. For the prevention or treatment of disease, the appropriate dosage may depend on the type of disease to be treated, the type of cells or recombinant receptors, the severity and course of the disease, whether the cells are administered for preventive or therapeutic purposes, previous therapy, the subject's clinical history and response to the cells, and the discretion of the attending physician. The compositions and cells are in some embodiments suitably administered to the subject at one time or over a series of treatments. In some embodiments, the cells are administered as part of a combination treatment, such as simultaneously with or sequentially with, in any order, another therapeutic intervention, such as an antibody or engineered cell or receptor or agent, such as a cytotoxic or therapeutic agent. The cells in some embodiments are co-administered with one or more additional therapeutic agents or in connection with another therapeutic intervention, either simultaneously or sequentially in any order. In some contexts, the cells are co-administered with another therapy sufficiently close in time such that the cell populations enhance the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, the cells are administered prior to the one or more additional therapeutic agents. In some embodiments, the cells are administered after the one or more additional therapeutic agents. In some embodiments, the one or more additional agents includes a cytokine, such as IL-2, for example, to enhance persistence. In some embodiments, the methods comprise administration of a chemotherapeutic agent. In certain embodiments, the modified cells of the disclosure (e.g., a modified cell comprising a CAR) may be administered to a subject in combination with an immune checkpoint antibody (e.g., an anti-PD1, anti-CTLA-4, or anti-PDL1 antibody). For example, the modified cell may be administered in combination with an antibody or antibody fragment targeting, for example, PD-1 (programmed death 1 protein). Examples of anti-PD-1 antibodies include, but are not limited to, pembrolizumab (KEYTRUDA®, formerly lambrolizumab, also known as MK-3475), and nivolumab (BMS-936558, MDX-1106, ONO-4538, OPDIVA®) or an antigen-binding fragment thereof. In certain embodiments, the modified cell may be administered in combination with an anti- PD-L1 antibody or antigen-binding fragment thereof. Examples of anti-PD-L1 antibodies include, but are not limited to, BMS-936559, MPDL3280A (TECENTRIQ®, Atezolizumab), and MEDI4736 (Durvalumab, Imfinzi). In certain embodiments, the modified cell may be administered in combination with an anti-CTLA-4 antibody or antigen-binding fragment thereof. An example of an anti-CTLA-4 antibody includes, but is not limited to, Ipilimumab (trade name Yervoy). Other types of immune checkpoint modulators may also be used including, but not limited to, small molecules, siRNA, miRNA, and CRISPR systems. Immune checkpoint modulators may be administered before, after, or concurrently with the modified cell comprising the CAR. In certain embodiments, combination treatment comprising an immune checkpoint modulator may increase the therapeutic efficacy of a therapy comprising a modified cell of the present disclosure. Following administration of the cells, the biological activity of the engineered cell populations in some embodiments is measured, e.g., by any of a number of known methods. Parameters to assess include specific binding of an engineered or natural T cell or other immune cell to antigen, in vivo, e.g., by imaging, or ex vivo, e.g., by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable method known in the art, such as cytotoxicity assays described in, for example, Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009); Herman et al. J. Immunological Methods, 285(1): 25-40 (2004); Kiesgen et al., Nat Protoc. (2021) 16(3):1331-1342; and Maldini et al., J Immunol Methods (2020) 484-485:112830. In certain embodiments, the biological activity of the cells is measured by assaying expression and / or secretion of one or more cytokines, such as CD 107a, IFNγ, IL-2, and TNF. In some aspects the biological activity is measured by assessing clinical outcome, such as reduction in tumor burden or load. In certain embodiments, the subject is provided a secondary treatment. Secondary treatments include but are not limited to chemotherapy, radiation, surgery, and medications. In some embodiments, the subject can be administered a conditioning therapy prior to CAR T cell therapy. In some embodiments, the conditioning therapy comprises administering an effective amount of cyclophosphamide to the subject. In some embodiments, the conditioning therapy comprises administering an effective amount of fludarabine to the subject. In preferred embodiments, the conditioning therapy comprises administering an effective amount of a combination of cyclophosphamide and fludarabine to the subject. Administration of a conditioning therapy prior to CAR T cell therapy may increase the efficacy of the CAR T cell therapy. Methods of conditioning patients for T cell therapy are described in U.S. Pat. No. 9,855,298, which is incorporated herein by reference in its entirety. In some embodiments, a specific dosage regimen of the present disclosure includes a lymphodepletion step prior to the administration of the modified T cells. In an exemplary embodiment, the lymphodepletion step includes administration of cyclophosphamide and / or fludarabine. Cells of the disclosure can be administered in dosages and routes and at times to be determined in appropriate pre-clinical and clinical experimentation and trials. Cell compositions may be administered multiple times at dosages within these ranges. Administration of the cells of the disclosure may be combined with other methods useful to treat the desired disease or condition as determined by those of skill in the art. It is known in the art that one of the adverse effects following infusion of CAR T cells is the onset of immune activation, known as cytokine release syndrome (CRS). CRS is immune activation resulting in elevated inflammatory cytokines. CRS is a known on-target toxicity, development of which likely correlates with efficacy. Clinical and laboratory measures range from mild CRS (constitutional symptoms and / or grade-2 organ toxicity) to severe CRS (sCRS; grade ≥3 organ toxicity, aggressive clinical intervention, and / or potentially life threatening). Clinical features include: high fever, malaise, fatigue, myalgia, nausea, anorexia, tachycardia / hypotension, capillary leak, cardiac dysfunction, renal impairment, hepatic failure, and disseminated intravascular coagulation. Dramatic elevations of cytokines including interferon-gamma, granulocyte macrophage colony-stimulating factor, IL-10, and IL-6 have been shown following CAR T-cell infusion. One CRS signature is elevation of cytokines including IL-6 (severe elevation), IFN-gamma, TNF-alpha (moderate), and IL-2 (mild). Elevations in clinically available markers of inflammation including ferritin and C-reactive protein (CRP) have also been observed to correlate with the CRS syndrome. The presence of CRS generally correlates with expansion and progressive immune activation of adoptively transferred cells. It has been demonstrated that the degree of CRS severity is dictated by disease burden at the time of infusion as patients with high tumor burden experience a more sCRS. Accordingly, the disclosure provides for, following the diagnosis of CRS, appropriate CRS management strategies to mitigate the physiological symptoms of uncontrolled inflammation without dampening the antitumor efficacy of the engineered cells (e.g., CAR T cells). CRS management strategies are known in the art. For example, systemic corticosteroids may be administered to rapidly reverse symptoms of sCRS (e.g., grade 3 CRS) without compromising initial antitumor response. In some embodiments, an anti-IL-6R antibody may be administered. An example of an anti- IL-6R antibody is the Food and Drug Administration-approved monoclonal antibody tocilizumab, also known as atlizumab (marketed as Actemra, or RoActemra). Tocilizumab is a humanized monoclonal antibody against the interleukin-6 receptor (IL-6R). Administration of tocilizumab has demonstrated near-immediate reversal of CRS. CRS is generally managed based on the severity of the observed syndrome and interventions are tailored as such. CRS management decisions may be based upon clinical signs and symptoms and response to interventions, not solely on laboratory values alone. Mild to moderate cases generally are treated with symptom management with fluid therapy, non-steroidal anti-inflammatory drug (NSAID) and antihistamines as needed for adequate symptom relief. More severe cases include patients with any degree of hemodynamic instability; with any hemodynamic instability, the administration of tocilizumab is recommended. The first-line management of CRS may be tocilizumab, in some embodiments, at the labeled dose of 8 mg / kg IV over 60 minutes (not to exceed 800 mg / dose); tocilizumab can be repeated Q8 hours. If suboptimal response to the first dose of tocilizumab, additional doses of tocilizumab may be considered. Tocilizumab can be administered alone or in combination with corticosteroid therapy. Patients with continued or progressive CRS symptoms, inadequate clinical improvement in 12-18 hours or poor response to tocilizumab, may be treated with high-dose corticosteroid therapy, generally hydrocortisone 100 mg IV or methylprednisolone 1-2 mg / kg. In patients with more severe hemodynamic instability or more severe respiratory symptoms, patients may be administered high- dose corticosteroid therapy early in the course of the CRS. CRS management guidance may be based on published standards (Lee et al. (2019) Biol Blood Marrow Transplant, doi.org / 10.1016 / j.bbmt.2018.12.758; Neelapu et al. (2018) Nat Rev Clin Oncology, 15:47; Teachey et al. (2016) Cancer Discov, 6(6):664-679). Features consistent with Macrophage Activation Syndrome (MAS) or Hemophagocytic lymphohistiocytosis (HLH) have been observed in patients treated with CAR-T therapy (Henter, 2007), coincident with clinical manifestations of the CRS. MAS appears to be a reaction to immune activation that occurs from the CRS, and should therefore be considered a manifestation of CRS. MAS is similar to HLH (also a reaction to immune stimulation). The clinical syndrome of MAS is characterized by high grade non-remitting fever, cytopenias affecting at least two of three lineages, and hepatosplenomegaly. It is associated with high serum ferritin, soluble interleukin-2 receptor, and triglycerides, and a decrease of circulating natural killer (NK) activity. In one aspect, the disclosure includes a method of treating cancer in a subject in need thereof, comprising administering to the subject any one of the modified immune or precursor cells disclosed herein. Yet another aspect of the disclosure includes a method of treating cancer in a subject in need thereof, comprising administering to the subject a modified immune or precursor cell generated by any one of the methods disclosed herein. Sources of Immune Cells In certain embodiments, a source of immune cells (e.g. T cells) is obtained from a subject for ex vivo manipulation and / or in vivo transduction. Sources of target cells for ex vivo manipulation may also include, e.g., autologous or heterologous donor blood, cord blood, or bone marrow. For example, the source of immune cells may be from the subject to be treated with the modified immune cells of the disclosure, e.g., the subject's blood, the subject's cord blood, or the subject's bone marrow. Non-limiting examples of subjects include humans, dogs, cats, mice, rats, and transgenic species thereof. Preferably, the subject is a human. Methods for in vivo transduction of immune cells for CAR expression are described, e.g., in Pfeiffer et al., EMBO Mol Med. (2018) 10(11):e9158; Weidner et al., Nat Protoc. (2021) 16(7):3210-3240; Frank et al., Blood Advances (2020) 4(22):5702-5715; Nawaz et al., Blood Cancer J. (2021) 11(6):119. Immune cells can be obtained from a number of sources, including blood, peripheral blood mononuclear cells, bone marrow, lymph node tissue, spleen tissue, umbilical cord, lymph, or lymphoid organs. Immune cells are cells of the immune system, such as cells of the innate or adaptive immunity, e.g., myeloid or lymphoid cells, including lymphocytes, typically T cells and / or NK cells. Other exemplary cells include stem cells, such as multipotent and pluripotent stem cells, including induced pluripotent stem cells (iPSCs). In some aspects, the cells are human cells. With reference to the subject to be treated, the cells may be allogeneic and / or autologous. The cells typically are primary cells, such as those isolated directly from a subject and / or isolated from a subject and frozen. In certain embodiments, the immune cell is a T cell, e.g., a CD8+ T cell (e.g., a CD8+ naive T cell, central memory T cell, or effector memory T cell), a CD4+ T cell, a natural killer T cell (NKT cells), a regulatory T cell (Treg), a stem cell memory T cell, a lymphoid progenitor cell, a hematopoietic stem cell, a natural killer cell (NK cell), a macrophage, or a dendritic cell. In some embodiments, the cells are monocytes or granulocytes, e.g., myeloid cells, macrophages, neutrophils, dendritic cells, mast cells, eosinophils, and / or basophils. In an embodiment, the target cell is an induced pluripotent stem (iPS) cell or a cell derived from an iPS cell, e.g., an iPS cell generated from a subject, manipulated to alter (e.g., induce a mutation in) or manipulate the expression of one or more target genes, and differentiated into, e.g., a T cell, e.g., a CD8+ T cell (e.g., a CD8+naive T cell, central memory T cell, or effector memory T cell), a CD4+T cell, a stem cell memory T cell, a lymphoid progenitor cell, or a hematopoietic stem cell. In some embodiments, the cells include one or more subsets of T cells or other cell types, such as whole T cell populations, CD4+ cells, CD8+ cells, and subpopulations thereof, such as those defined by function, activation state, maturity, potential for differentiation, expansion, recirculation, localization, and / or persistence capacities, antigen-specificity, type of antigen receptor, presence in a particular organ or compartment, marker or cytokine secretion profile, and / or degree of differentiation. Among the sub-types and subpopulations of T cells and / or of CD4+and / or of CD8+T cells are naive T (TN) cells, effector T cells (TEFF), memory T cells and sub-types thereof, such as stem cell memory T (TSCM), central memory T (TCM), effector memory T (TEM), or terminally differentiated effector memory T cells, tumor-infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosa-associated invariant T (MAIT) cells, naturally occurring and adaptive regulatory T (Treg) cells, helper T cells, such as TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, alpha / beta T cells, and delta / gamma T cells. In certain embodiments, any number of T cell lines available in the art, may be used. In some embodiments, the methods include isolating immune cells from the subject, preparing, processing, culturing, and / or engineering them. In some embodiments, preparation of the engineered cells includes one or more culture and / or preparation steps. The cells for engineering as described may be isolated from a sample, such as a biological sample, e.g., one obtained from or derived from a subject. In some embodiments, the subject from which the cell is isolated is one having the disease or condition or in need of a cell therapy or to which cell therapy will be administered. The subject in some embodiments is a human in need of a particular therapeutic intervention, such as the adoptive cell therapy for which cells are being isolated, processed, and / or engineered. Accordingly, the cells in some embodiments are primary cells, e.g., primary human cells. The samples include tissue, fluid, and other samples taken directly from the subject, as well as samples resulting from one or more processing steps, such as separation, centrifugation, genetic engineering (e.g. transfection with a nucleic acid molecule of the disclosure, optionally in the presence of an enzyme capable of performing genomic integration of the nucleic acid molecule), washing, and / or incubation. The biological sample can be a sample obtained directly from a biological source or a sample that is processed. Biological samples include, but are not limited to, body fluids, such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine and sweat, tissue and organ samples, including processed samples derived therefrom. In some aspects, the sample from which the cells are derived or isolated is blood or a blood- derived sample, or is or is derived from an apheresis or leukapheresis product. Exemplary samples include whole blood, peripheral blood mononuclear cells (PBMCs), leukocytes, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut associated lymphoid tissue, mucosa associated lymphoid tissue, spleen, other lymphoid tissues, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testes, ovaries, tonsil, or other organ, and / or cells derived therefrom. Samples include, in the context of cell therapy, e.g., adoptive cell therapy, samples from autologous and allogeneic sources. In some embodiments, the cells are derived from cell lines, e.g., T cell lines. The cells in some embodiments are obtained from a xenogeneic source, for example, from mouse, rat, non- human primate, and pig. In some embodiments, isolation of the cells includes one or more preparation and / or non-affinity based cell separation steps. In some examples, cells are washed, centrifuged, and / or incubated in the presence of one or more reagents, for example, to remove unwanted components, enrich for desired components, lyse or remove cells sensitive to particular reagents. In some examples, cells are separated based on one or more property, such as density, adherent properties, size, sensitivity and / or resistance to particular components. In some examples, cells from the circulating blood of a subject are obtained, e.g., by apheresis or leukapheresis. The samples, in some aspects, contain lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and / or platelets, and in some aspects contains cells other than red blood cells and platelets. In some embodiments, the blood cells collected from the subject are washed, e.g., to remove the plasma fraction and to place the cells in an appropriate buffer or media for subsequent processing steps. In some embodiments, the cells are washed with phosphate buffered saline (PBS). In some aspects, a washing step is accomplished by tangential flow filtration (TFF) according to the manufacturer's instructions. In some embodiments, the cells are resuspended in a variety of biocompatible buffers after washing. In certain embodiments, components of a blood cell sample are removed and the cells directly resuspended in culture media. In some embodiments, the methods include density-based cell separation methods, such as the preparation of white blood cells from peripheral blood by lysing the red blood cells and centrifugation through a Percoll or Ficoll gradient. In one embodiment, immune cells are obtained from the circulating blood of an individual by apheresis or leukapheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. The cells collected by apheresis may be washed to remove the plasma fraction and to place the cells in an appropriate buffer or media, such as phosphate buffered saline (PBS) or wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations, for subsequent processing steps. After washing, the cells may be resuspended in a variety of biocompatible buffers, such as, for example, Ca-free, Mg-free PBS. Alternatively, the undesirable components of the apheresis sample may be removed and the cells directly resuspended in culture media. In some embodiments, the isolation methods include the separation of different cell types based on the expression or presence in the cell of one or more specific molecules, such as surface markers, e.g., surface proteins, intracellular markers, or nucleic acid. In some embodiments, any known method for separation based on such markers may be used. In some embodiments, the separation is affinity- or immunoaffinity-based separation. For example, the isolation in some aspects includes separation of cells and cell populations based on the cells' expression or expression level of one or more markers, typically cell surface markers, for example, by incubation with an antibody or binding partner that specifically binds to such markers, followed generally by washing steps and separation of cells having bound the antibody or binding partner, from those cells having not bound to the antibody or binding partner. Such separation steps can be based on positive selection, in which the cells having bound the reagents are retained for further use, and / or negative selection, in which the cells having not bound to the antibody or binding partner are retained. In some examples, both fractions are retained for further use. In some aspects, negative selection can be particularly useful where no antibody is available that specifically identifies a cell type in a heterogeneous population, such that separation is best carried out based on markers expressed by cells other than the desired population. The separation need not result in 100% enrichment or removal of a particular cell population or cells expressing a particular marker. For example, positive selection of or enrichment for cells of a particular type, such as those expressing a marker, refers to increasing the number or percentage of such cells, but need not result in a complete absence of cells not expressing the marker. Likewise, negative selection, removal, or depletion of cells of a particular type, such as those expressing a marker, refers to decreasing the number or percentage of such cells, but need not result in a complete removal of all such cells. In some examples, multiple rounds of separation steps are carried out, where the positively or negatively selected fraction from one step is subjected to another separation step, such as a subsequent positive or negative selection. In some examples, a single separation step can deplete cells expressing multiple markers simultaneously, such as by incubating cells with a plurality of antibodies or binding partners, each specific for a marker targeted for negative selection. Likewise, multiple cell types can simultaneously be positively selected by incubating cells with a plurality of antibodies or binding partners expressed on the various cell types. Expansion of Immune Cells Whether prior to or after modification of cells to express a CAR of a nucleic acid molecule of the disclosure, the cells can be activated and expanded in number using methods as described, for example, in U.S. Pat. Nos. 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869; 7,232,566; 7,175,843; 5,883,223; 6,905,874; 6,797,514; 6,867,041; and U.S. Publication No. 20060121005. For example, the T cells of the disclosure may be expanded by contact with a surface having attached thereto an agent that stimulates a CD3 / TCR complex associated signal and a ligand that stimulates a co-stimulatory molecule on the surface of the T cells. In particular, T cell populations may be stimulated by contact with an anti-CD3 antibody, or antigen-binding fragment thereof, or an anti-CD2 antibody immobilized on a surface, or by contact with a protein kinase C activator (e.g., bryostatin) in conjunction with a calcium ionophore. For co-stimulation of an accessory molecule on the surface of the T cells, a ligand that binds the accessory molecule is used. For example, T cells can be contacted with an anti-CD3 antibody and an anti-CD28 antibody, under conditions appropriate for stimulating proliferation of the T cells. Examples of an anti-CD28 antibody include 9.3, B-T3, XR- CD28 (Diaclone, Besancon, France) and these can be used in the disclosure, as can other methods and reagents known in the art (see, e.g., ten Berge et al., Transplant Proc. (1998) 30(8): 3975-3977; Haanen et al., J. Exp. Med. (1999) 190(9): 1319-1328; and Garland et al., J. Immunol. Methods (1999) 227(1-2): 53-63). Expanding T cells by the methods disclosed herein can be multiplied by about 10 fold, 20 fold, 30 fold, 40 fold, 50 fold, 60 fold, 70 fold, 80 fold, 90 fold, 100 fold, 200 fold, 300 fold, 400 fold, 500 fold, 600 fold, 700 fold, 800 fold, 900 fold, 1000 fold, 2000 fold, 3000 fold, 4000 fold, 5000 fold, 6000 fold, 7000 fold, 8000 fold, 9000 fold, 10,000 fold, 100,000 fold, 1,000,000 fold, 10,000,000 fold, or greater, and any and all whole or partial integers therebetween. In one embodiment, the T cells expand in the range of about 20 fold to about 50 fold. Following culturing, the T cells can be incubated in cell medium in a culture apparatus for a period of time or until the cells reach confluency or high cell density for optimal passage before passing the cells to another culture apparatus. The culturing apparatus can be of any culture apparatus commonly used for culturing cells in vitro. Preferably, the level of confluence is 70% or greater before passing the cells to another culture apparatus. More preferably, the level of confluence is 90% or greater. A period of time can be any time suitable for the culture of cells in vitro. The T cell medium may be replaced during the culture of the T cells at any time. Preferably, the T cell medium is replaced about every 2 to 3 days. The T cells are then harvested from the culture apparatus whereupon the T cells can be used immediately or cryopreserved to be stored for use at a later time. In one embodiment, the disclosure includes cryopreserving the expanded T cells. The cryopreserved T cells are thawed prior to introducing nucleic acids into the T cell. In another embodiment, the method comprises isolating T cells and expanding the T cells. In another embodiment, the disclosure further comprises cryopreserving the T cells prior to expansion. In yet another embodiment, the cryopreserved T cells are thawed for electroporation with the RNA encoding the chimeric membrane protein. Another procedure for ex vivo expansion cells is described in U.S. Pat. No. 5,199,942 (incorporated herein by reference). Expansion, such as described in U.S. Pat. No.5,199,942 can be an alternative or in addition to other methods of expansion described herein. Briefly, ex vivo culture and expansion of T cells comprises the addition to the cellular growth factors, such as those described in U.S. Pat. No.5,199,942, or other factors, such as flt3-L, IL-1, IL-3 and c-kit ligand. In one embodiment, expanding the T cells comprises culturing the T cells with a factor selected from the group consisting of flt3-L, IL-1, IL-3 and c-kit ligand. The medium used to culture the T cells may include an agent that can co-stimulate the T cells. For example, an agent that can stimulate CD3 is an antibody to CD3, and an agent that can stimulate CD28 is an antibody to CD28. A cell isolated by the methods disclosed herein can be expanded approximately 10 fold, 20 fold, 30 fold, 40 fold, 50 fold, 60 fold, 70 fold, 80 fold, 90 fold, 100 fold, 200 fold, 300 fold, 400 fold, 500 fold, 600 fold, 700 fold, 800 fold, 900 fold, 1000 fold, 2000 fold, 3000 fold, 4000 fold, 5000 fold, 6000 fold, 7000 fold, 8000 fold, 9000 fold, 10,000 fold, 100,000 fold, 1,000,000 fold, 10,000,000 fold, or greater. In one embodiment, the T cells expand in the range of about 20 fold to about 50 fold, or more. In one embodiment, human T regulatory cells are expanded via anti-CD3 antibody coated KT64.86 artificial antigen presenting cells (aAPCs). Methods for expanding and activating T cells can be found in U.S. Pat. Nos.7,754,482, 8,722,400, and 9,555,105, contents of which are incorporated herein in their entirety. In one embodiment, the method of expanding the T cells can further comprise isolating the expanded T cells for further applications. In another embodiment, the method of expanding can further comprise a subsequent electroporation of the expanded T cells followed by culturing. The subsequent electroporation may include introducing a nucleic acid encoding an agent, such as a transducing the expanded T cells, transfecting the expanded T cells, or electroporating the expanded T cells with a nucleic acid, into the expanded population of T cells, wherein the agent further stimulates the T cell. The agent may stimulate the T cells, such as by stimulating further expansion, effector function, or another T cell function. Pharmaceutical Compositions and Formulations Also provided are populations of immune cells of the disclosure, compositions containing such cells and / or enriched for such cells, such as in which cells expressing a CAR and / or nucleic acid molecule-encoded shRNAs make up at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more of the total cells in the composition or cells of a certain type such as T cells or CD8+ or CD4+ cells. Among the compositions are pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy. Also provided are therapeutic methods for administering the cells and compositions to subjects, e.g., patients. Also provided are compositions including the cells for administration, including pharmaceutical compositions and formulations, such as unit dose form compositions including the number of cells for administration in a given dose or fraction thereof. The pharmaceutical compositions and formulations generally include one or more optional pharmaceutically acceptable carrier or excipient. In some embodiments, the composition includes at least one additional therapeutic agent. The term “pharmaceutical formulation” or “pharmaceutical composition” refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. A “pharmaceutically acceptable carrier” refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, which is nontoxic to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, excipient, stabilizer, or preservative. In some aspects, the choice of carrier is determined in part by the particular cell and / or by the method of administration. Accordingly, there are a variety of suitable formulations. For example, the pharmaceutical composition can contain preservatives. Suitable preservatives may include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some aspects, a mixture of two or more preservatives is used. The preservative or mixtures thereof are typically present in an amount of about 0.0001% to about 2% by weight of the total composition. Carriers are described, e.g., by Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed, and include, but are not limited to: buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g. Zn- protein complexes); and / or non-ionic surfactants such as polyethylene glycol (PEG). Buffering agents in some aspects are included in the compositions. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more buffering agents is used. The buffering agent or mixtures thereof are typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005). The formulations can include aqueous solutions. The formulation or composition may also contain more than one active ingredient useful for the particular indication, disease, or condition being treated with the cells, preferably those with activities complementary to the cells, where the respective activities do not adversely affect one another. Such active ingredients are suitably present in combination in amounts that are effective for the purpose intended. Thus, in some embodiments, the pharmaceutical composition further includes other pharmaceutically active agents or drugs, such as chemotherapeutic agents, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, and / or vincristine. The pharmaceutical composition in some embodiments contains the cells in amounts effective to treat or prevent the disease or condition, such as a therapeutically effective or prophylactically effective amount. Therapeutic or prophylactic efficacy in some embodiments is monitored by periodic assessment of treated subjects. The desired dosage can be delivered by a single bolus administration of the cells, by multiple bolus administrations of the cells, or by continuous infusion administration of the cells. Formulations include those for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. In some embodiments, the cell populations are administered parenterally. The term “parenteral,” as used herein, includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. In some embodiments, the cells are administered to the subject using peripheral systemic delivery by intravenous, intraperitoneal, or subcutaneous injection. Compositions in some embodiments are provided as sterile liquid preparations, e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may in some aspects be buffered to a selected pH. Liquid preparations are normally easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions are somewhat more convenient to administer, especially by injection. Viscous compositions, on the other hand, can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyoi (for example, glycerol, propylene glycol, liquid polyethylene glycol) and suitable mixtures thereof. Sterile injectable solutions can be prepared by incorporating the cells in a solvent, such as in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose, dextrose, or the like. The compositions can contain auxiliary substances such as wetting, dispersing, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity enhancing additives, preservatives, flavoring agents, and / or colors, depending upon the route of administration and the preparation desired. Standard texts may in some aspects be consulted to prepare suitable preparations. Various additives which enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers, can be added. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, and sorbic acid. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin. The formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, e.g., by filtration through sterile filtration membranes. In embodiments, there is provided a transgenic organism that may comprise cells which have been transformed with the nucleic acid constructs and / or have received the engineered cells of the present disclosure. In embodiments, the organism may be a mammal or an insect. When the organism is a mammal, the organism may include, but is not limited to, a mouse, a rat, a monkey, a dog, a rabbit and the like. When the organism is an insect, the organism may include, but is not limited to, a fruit fly, a mosquito, a bollworm and the like. Kits Compositions of the disclosure can be included in a container, kit, pack, or dispenser together with instructions for administration. Also provided herein are kits comprising: i) any of the aforementioned gene transfer constructs of this invention, and / or any of the aforementioned cells of this invention and ii) a container. In certain embodiments, the kits further comprise instructions for the use thereof. In certain embodiments, any of the aforementioned kits can further comprise a recombinant DNA construct comprising a CAR-encoding sequence, one or more shRNA-encoding sequences, and / or terminal repeat sequences. Optionally, any of the aforementioned kits can further comprise a nucleic acid sequence that encodes a transposase. A kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition. The practice of the present disclosure employs, unless otherwise indicated, conventional techniques of chemistry, molecular biology, microbiology, recombinant DNA, genetics, immunology, cell biology, cell culture and transgenic biology, which are within the skill of the art. See, e.g., Maniatis et al., 1982, Molecular Cloning (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Sambrook et al., 1989, Molecular Cloning, 2nd Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Sambrook and Russell, 2001, Molecular Cloning, 3rd Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Ausubel et al., 1992), Current Protocols in Molecular Biology (John Wiley & Sons, including periodic updates); Glover, 1985, DNA Cloning (IRL Press, Oxford); Anand, 1992; Guthrie and Fink, 1991; Harlow and Lane, 1988, Antibodies, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Jakoby and Pastan, 1979; Nucleic Acid Hybridization (B. D. Hames & S. J. Higgins eds.1984); Transcription And Translation (B. D. Hames & S. J. Higgins eds.1984); Culture Of Animal Cells (R. I. Freshney, Alan R. Liss, Inc., 1987); Immobilized Cells And Enzymes (IRL Press, 1986); B. Perbal, A Practical Guide To Molecular Cloning (1984); the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.); Gene Transfer Vectors For Mammalian Cells (J. H. Miller and M. P. Calos eds., 1987, Cold Spring Harbor Laboratory); Methods In Enzymology, Vols. 154 and 155 (Wu et al. eds.), Immunochemical Methods In Cell And Molecular Biology (Mayer and Walker, eds., Academic Press, London, 1987); Handbook Of Experimental Immunology, Volumes I- IV (D. M. Weir and C. C. Blackwell, eds., 1986); Riott, Essential Immunology, 6th Edition, Blackwell Scientific Publications, Oxford, 1988; Hogan et al., Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986); Westerfield, M., The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio), (4th Ed., Univ. of Oregon Press, Eugene, 2000). Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Reference will now be made in detail to exemplary embodiments of the disclosure. While the disclosure will be described in conjunction with the exemplary embodiments, it will be understood that it is not intended to limit the disclosure to those embodiments. To the contrary, it is intended to cover alternatives, modifications, and equivalents as may be included within the spirit and scope of the disclosure as defined by the appended claims. Standard techniques well known in the art or the techniques specifically described below were utilized. EXAMPLES Example 1: Materials and Methods Nucleic Acid Constructs Silencer select siRNA oligonucleotides were purchased from Thermo Fisher. Plasmid DNA was purchased from GenScript. Nanoplasmid DNA was purchased from Nature Technology / Aldevron. The SaliogaseB enzyme was produced from in vitro transcription (IVT) of a T7-driven vector containing xenopus globin 5’ and 3’ UTRs with 5-methyl- pseudo-U modification and a synthetic 34 polyA tail (TriLink). Cell Lines HH cells were purchased from ATCC (CRL-2105), cultured in RPMI 1640 Medium (ATCC modified, Gibco, #A1049101) with 10% heat inactivated fetal bovine serum (Gibco #10438026), and reseeded to a density of 2 x 105cells / mL every 3-4 days. Transfections HH cells were transfected with the SE Primary Cell 4D-Nucleofector™ X Kit L (Lonza, # V4XC-1024). Transfections were performed in 100 µL reactions with 1 x 106cells per condition with 2 µg of donor DNA and 10 µg SaliogaseB mRNA (1:5 ratio) using program CL-120. Flow Cytometry detection of shRNA activity and CAR Expression 200 µL of cell suspension was stained with zombie violet dye (Biolegend, #77477) for viability, FITC anti-human HLA-A,B,C (Biolgened, #311404) for MHC class I detection, and APC labeled CD19 protein (AcroBio, #CD19HA2H9) for CD19-CAR detection following manufacturers’ protocols. Analysis was performed with a Beckman Coulter CytoFLEX S flow cytometer. Example 2: CAR-T Cell Design for Allogeneic Therapy Design and use of engineered T cells expressing chimeric antigen receptors (CARs) in both autologous cell therapies and in allogeneic cell therapies have been widely described. CAR-T cells have been designed to target specific cell-surface antigens (e.g., tumor antigens / markers) and elicit an immune response upon antigen detection. CAR-T cells can be engineered for general immunotherapies, to treat cancer, and for other purposes. CAR-T cells can be engineered for autologous cell therapies or allogeneic cell therapies. Autologous CAR-T therapies introduce a CAR to cells derived from a patient's own sample (e.g., a blood sample): T cells are isolated from a patient sample, are then genetically engineered to perform a specific function, and are administered back into the same patient's body to perform that function. In contrast, in allogeneic CAR-T cell therapies, an initial sample is taken from a donor, T cells are isolated from such sample and are then engineered to express a CAR, the CAR-expressing cell population is expanded, and the engineered CAR-T cells are then administered to patients other than the original donor. Allogeneic CAR-T cell therapies offer certain advantages over autologous cell therapies, as allogeneic therapies allow for widespread and simplified distribution of such cell therapies, provided that such cells are not rejected by the non-self recipient after administration. Design and implementation of an allogeneic CAR-T cell therapy was first envisioned for CD19+CAR-T cells, involving knockdown of MHC class I, MHC class II and / or TCRα proteins, coupled with knock-in of an anti-CD19 CAR and optionally knock-in of additional cell surface receptors (e.g., CD47; FIG.1). To perform knockdown of MHC class I, MHC class II and / or TCRα proteins, shRNAs targeting T cell receptor alpha constant (TRAC, as a TCRα target, noting that CD3ζ may also or alternatively be targeted; FIG. 2), beta 2 microglobulin (B2M, an MHC class I target), transporter 1, ATP binding cassette subfamily B member (TAP1, an MHC class I target; FIG.3), transporter 2, ATP binding cassette subfamily B member (TAP2, an MHC class I target), and Class II Major Histocompatibility Complex Transactivator (CIITA, an MHC class II target) were designed. MHC I and II knockdowns are performed to prevent recognition of allogeneic cells by circulating CD8+T-cells, thereby allowing such allogeneic cells to persist. The T-cell receptor complex (specifically, TCRα) is also knocked down to avoid T-cell receptor (TCR)-MHC complex formation between allogeneic T-cells and antigen-presenting cells (APCs; FIG.2), thereby allowing exogenous CAR-expressing cells to function. Knock-in of CD47 can also be performed, to prevent allogeneic T-cells from being destroyed by circulating macrophages. Example 3: Selection of shRNA Screening Cell Line(s) and Transfection To identify a preferred cell line for performance of shRNA screening, HH cells (a mature T cell line derived from peripheral blood of a patient with aggressive cutaneous T cell leukemia / lymphoma) were assessed as a potential target cell line for shRNAs, to be used in screening. Flow cytometry was used to identify high cell surface levels of both MHC class I and MHC class II proteins in HH cells (FIG. 4). Levels of beta 2 microglobulin (B2M) mRNA in HH cells were nearly as high as GAPDH control mRNA levels, as measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR), all of which contributed to HH cells being identified as a good shRNA screening cell line (particularly for assessment of B2M knockdown by B2M-targeting shRNAs). HH cells were shown to transfect well (as demonstrated by intracellular accumulation of a transfected GFP marker) and also showed robust cellular viability post- transfection (delivery and viability levels exceeding 80% of cells in the transfected population were respectively observed). HH cells were therefore assessed as showing good transfection and cell growth (FIG.4). Example 4: CAR - shRNA Construct Design Various formats of shRNA constructs were initially designed, made and assessed for activity (FIG.5). These initial constructs exhibited only a modest prevalence of CAR delivery (roughly 25- 60%), with little to no shRNA-mediated knockdown of target gene sequences. The initial constructs were therefore redesigned, with focus upon six distinct shRNA regions of such test constructs: (1) orientation of target / seed strands – switching of strand locations in shRNAs (assessed with "P217" and "P218" constructs herein); (2) proximity of shRNA sequence to the U6 promoter – introducing spacer sequences (e.g., 10bp spacer sequence, as assessed with constructs "P219" and "P220"); (3) adding a polyA sequence; (4) making pri-miRNA constructs, if minimal shRNA constructs were ineffective; (5) using an alternative promoter (H1 RNA polymerase III promoter rather than U6, as was used for "P227" and "P228" constructs); and (6) including a larger loop size (e.g., a 12 nt loop, as was used in "P229" and "P230" constructs) (FIG.6). Among the redesigned constructs, the "P220" U6 promoter-shRNA linker addition construct (in which a 10 bp linker was inserted between the U6 promoter and the shRNA-encoding sequence) was most effective. The P220 construct shRNA successfully knocked down targeted MHC class I expression, as observed both by lower mean fluorescence intensity at the cell surface in live cells (FIG.7A) and by MHC class I levels as assessed by qRT-PCR (FIG.7B). The P220 shRNA construct (containing a beta 2 microglobulin (B2M)-targeting shRNA) was also observed to effectively and significantly reduce MHC class I expression levels, with peak shRNA-mediated knockdown of MHC class 1 observed at day 7 (FIG.8, bottom right, where MHC class I levels were plotted versus side scatter area (SSC-A)). Indeed, dramatic reduction of MHC class I levels was observed between day three (D3) and day seven (D7) when the P220 shRNA construct was administered (FIG.9). The P220 shRNA construct was also effective at knocking down MHC class I levels in Jurkat cells at day 7 and day 11 post-administration, as compared to all other shRNA constructs tested (FIG.10). Robust shRNA-mediated knockdown (knockdown levels of greater than 60%) of two different complexes (MHC class I and TCR) was observed for shRNAs respectively targeting beta 2 microglobulin (B2M) and CD3ζ (FIGs.11A and 11B), with such effects observed across different tested cell lines in vitro (FIG.11C).21B2M-targeting shRNA-encoding sequences were designed: (1) A P287 CD19-CAR B2M.2 shRNA P220 construct (SEQ ID NO: 1), having a 10 bp P220 linker and a 100 bp interpromoter spacer (FIG.12); (2) A P288 B2M.2 shRNA P220 CD19-CAR construct (SEQ ID NO: 2), having a 10 bp P220 linker and a 100 bp interpromoter spacer (FIG.13); and (3) A P289 B2M.2 shRNA P220 reverse CD19-CAR construct (SEQ ID NO: 3), having a 10 bp P220 linker and a 100 bp interpromoter spacer (FIG.14). The above P287, P288 and P289 constructs each exhibited robust co-expression of both CD19-CAR and B2M-targeting shRNAs, with B2M-targeting shRNA expression assessed via measurement of MHC class I knockdown in treated cells (FIGs.15A and 15B). The B2M shRNA of each of the P287, P288 and P289 constructs was expressed and active, as demonstrated by robust (60-90%) levels of MHC class I knockdown observed for each such B2M-targeting shRNA- containing construct, in both Jurkat cells and T cells, as compared to the CD19-CAR-only negative control construct (FIG.15B). An exemplary manufacturing process for allogeneic CAR cells was also designed, and cell sorting experiments demonstrated successful knock-in and expression of CAR (at approx.90% levels) in cells treated with either a CD19-CAR only construct or with a CD19- CAR + B2M-targeting shRNA co-expression construct (FIG. 16). Meanwhile, expression of the B2M-targeting shRNA of each such co-expression construct was also demonstrated, as observed by efficacy in knocking down MHC class I at > 70% levels, as compared to the CD19-CAR only control construct (FIG.16). Further constructs that integrate P220-type linkers between promoters and CD3ζ-targeting shRNA-encoding sequences were designed: (4) A P331 CD19-CAR CD3z shRNA1 construct (SEQ ID NO: 4), having an 11 bp P220 linker and a 100 bp interpromoter spacer (FIG.17); (5) A P332 CD19-CAR CD3z shRNA2 construct (SEQ ID NO: 5), having an 11 bp P220 linker and a 100 bp interpromoter spacer (FIG.18) A single construct multiplexing shRNA-encoding sequences (for B2M and CIITA) was also designed (FIG. 19). Similarly, a construct that multiplexes shRNA-encoding sequences for B2M and CD3ζ (CD3z) was also made (FIG.20).An exemplary construct sequence showing the junction of U6 promoter with linker and shRNA-encoding sequences was also considered in detail (FIG. 21A), as a basis for preparation of a series of seventeen newly modified shRNAs. Design of five of the seventeen new shRNAs was based upon internal hits from HH and / or Jurkat cells (H1 promoter; Hit P228 – Target; Hits OL54 and OL64 – Reverse strands; Hit P218; FIG. 21B), while the remaining twelve shRNAs carried new modifications based upon the literature (FIG.21B). A two- dimensional schematic of an exemplary B2M-targeting shRNA cassette of the disclosure (SEQ ID NO: 80), which labels respective spacer / leader, target (double-stranded region), loop and terminal domains, served as a reference for the location of such newly tested modifications (FIG.21C). Six of the seventeen new shRNA constructs screened in T cells showed similar or increased knockdown of MHC class I as the parent P220 expression cassette shRNA (> 65% knockdown of MHC class I; FIG. 22A), while measurable benefits to knockdown levels were observed for shRNAs including new loop sequences and a 19A bulge (FIG. 22A). Specific shRNA expression constructs that showed equal or greater MHC class I knockdown in tested T cells than the P220 control included: a shRNA cassette having a 7 bp loop (70% knockdown); a shRNA cassette having a 9 bp loop (71% knockdown); a shRNA cassette having a 25 nt target (69% knockdown); a shRNA cassette having a 19A bulge (72% knockdown); a shRNA cassette having a downstream SV40 (66% knockdown); and a shRNA cassette having a downstream cPPT / CTS configuration (65% knockdown)(FIG.22A). Cell sorting experiments further validated such findings. When cells treated with the B2M shRNA were assessed for B2M shRNA-mediated knockdown of MHC class I, the tested B2M-targeting shRNA exhibited significant reduction of MHC class I levels by cell sorting analysis, relative to an appropriate non-treated control population of cells (FIG.22B). Example 5: Additional shRNA Constructs Additional constructs are designed, e.g., with variable U6 promoter distances, alternative stems and loops, and combined features, initially to target TRAC and / or B2M with construct- encoded shRNAs, and optionally involving other knockdown targets. Further constructs are also made that incorporate multiple target sites, either within a single gene / transcript or across multiple genes / transcripts. Example 6: Allogeneic CAR-T Cell Treatments In an exemplary treatment, immune cells are harvested from a donor and are then manipulated to knock-in an anti-CD19 CAR, while also performing a genetic modification of such cells to stably express one or more shRNAs targeting one or more of the following for shRNA- mediated knockdown: TCR (optionally TCRα, MHC I and / or MHC II). Knock-in of an additional cell surface receptor (e.g., CD47) is optionally also performed. A subject having a hematological cancer (e.g., leukemia, lymphoma, multiple myeloma, etc.) is identified, and is administered an effective amount of the genetically manipulated, CAR-expressing immune cells described above as an allogeneic therapy, thereby treating the hematological cancer in the subject.

Claims

We Claim:

1. A nucleic acid molecule comprising: (a) a type III RNA polymerase III promoter operably linked to a sequence encoding for a short hairpin RNA (shRNA), wherein the shRNA is capable of annealing and reducing expression of a messenger RNA (mRNA) selected from the group consisting of a MHC class I gene mRNA, a MHC class II gene mRNA and a TCRα chain mRNA; and (b) two terminal repeat sequences capable of recognition by an enzyme capable of performing targeted genomic integration of the nucleic acid molecule, wherein the enzyme is derived from Bombyx mori, Xenopus tropicalis, Trichoplusia ni, Myotis lucifugus, Rhinolophus ferrumequinum, Rousettus aegyptiacus, Phyllostomus discolor, Myotis myotis, Pteropus vampyrus, Pipistrellus kuhlii, troglodytes, Molossus molossus, or Homo sapiens.

2. The nucleic acid molecule of claim 1, further comprising a sequence encoding for a chimeric antigen receptor (CAR), optionally wherein the chimeric antigen receptor is an anti-CD19 CAR, optionally wherein the anti-CD19 CAR is operably linked to a eukaryotic translation elongation factor 1 α (EF-1α) promoter.

3. The nucleic acid molecule of claim 2, wherein the CAR comprises an anti-CD19 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence.

4. The nucleic acid molecule of any one of the preceding claims, wherein the nucleic acid molecule comprises two terminal repeat sequences.

5. The nucleic acid molecule of any one of the preceding claims comprising an ordered series of sequences selected from the group consisting of: (a) terminal repeat sequence – EF1α promoter operably linked to a CAR-encoding sequence – U6 promoter operably linked to a shRNA – terminal repeat sequence; (b) terminal repeat sequence – U6 promoter operably linked to a shRNA – EF1α promoter operably linked to a CAR-encoding sequence – terminal repeat sequence, wherein the shRNA and the CAR-encoding sequence are transcribed in the same direction;(c) terminal repeat sequence – U6 promoter operably linked to a shRNA – EF1α promoter operably linked to a CAR-encoding sequence – terminal repeat sequence, wherein the shRNA and the CAR-encoding sequence are transcribed in opposite directions; and (d) terminal repeat sequence – EF1α promoter operably linked to a CAR-encoding sequence – U6 promoter operably linked to two or more shRNA sequences – terminal repeat sequence.

6. The nucleic acid molecule of claim 5, wherein the CAR-encoding sequence encodes for an anti-CD19 CAR, optionally wherein the CAR comprises an anti-CD19 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence.

7. The nucleic acid molecule of claim 5, wherein the CAR-encoding sequence encodes for an anti-CD20 CAR, optionally wherein the CAR comprises an anti-CD20 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence.

8. The nucleic acid molecule of claim 5, wherein the CAR-encoding sequence encodes for an anti-CD22 CAR, optionally wherein the CAR comprises an anti-CD22 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB signaling domain sequence and a CD3 zeta signaling domain sequence.

9. The nucleic acid molecule of claim 5, wherein the CAR-encoding sequence encodes for an anti-B7 H3 CAR, optionally wherein the CAR comprises an anti-B7 H3 FMC light chain, a CD8 hinge and transmembrane domain, a 4-1BB or CD28 signaling domain sequence and a CD3 zeta signaling domain sequence.

10. The nucleic acid molecule of claim 5, wherein the CAR-encoding sequence encodes for an anti-human epidermal growth factor receptor 2 (Her2) CAR, optionally wherein the CAR comprises an anti-Her2 FMC light chain, a CD8 hinge and transmembrane domain, a CD28 signaling domain sequence and a CD3 zeta signaling domain sequence.

11. The nucleic acid molecule of claim 5, wherein the shRNA is a Beta-2-microglobulin- targeting shRNA.

12. The nucleic acid molecule of claim 5, wherein the shRNA is a T cell receptor alpha constant (TRAC)-targeting shRNA.

13. The nucleic acid molecule of claim 5, wherein the shRNA is a Class II Major Histocompatibility Complex Transactivator (CIITA)-targeting shRNA.

14. The nucleic acid molecule of claim 5, wherein the shRNA is a transporter 1, ATP binding cassette subfamily B member (TAP1)-targeting shRNA.

15. The nucleic acid molecule of claim 5, wherein the shRNA is a transporter 2, ATP binding cassette subfamily B member (TAP2)-targeting shRNA.

16. The nucleic acid molecule of claim 5, wherein the shRNA is a (CD3ζ)-targeting shRNA.

17. The nucleic acid molecule of claim 5 comprising a sequence selected from the group consisting of SEQ ID NOs: 1-5, 12-15, 57, 58, and 76-80.

18. The nucleic acid molecule of any one of the preceding claims comprising a linker sequence between sequences encoding for shRNAs, wherein the linker sequence is selected from the group consisting of SEQ ID NOs: 59-65.

19. The nucleic acid molecule of any one of the preceding claims, wherein the enzyme capable of performing targeted genomic integration is selected from the group consisting of Tn1, Tn2, Tn3, Tn5, Tn7, Tn9, Tn10, Tn552, Tn903, Tn1000 / Gamma-delta, Tn / O, tnsA, tnsB, tnsC, tniQ, IS10, ISS, IS911, Minos, Sleeping beauty, piggyBac, Tol2, Mos1, Himar1, Hermes, Tol2, Minos, Tel, P- element, MuA, Ty1, Chapaev, transib, Tc1 / mariner, and Tc3 donor DNA system.

20. The nucleic acid molecule of any one of the preceding claims, wherein the enzyme capable of performing targeted genomic integration comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 66, 69-75, and variants thereof.

21. The nucleic acid molecule of any one of the preceding claims, wherein the two repeat sequences comprisea Myotis lucifugus mobile element enzyme (MLT) left terminal end (SEQ ID NO: 67), or a sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto; and / or a MLT right terminal end (SEQ ID NO: 68), or a sequence having at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto.

22. The nucleic acid molecule of any one of the preceding claims further comprising a rabbit β- globin polyadenylation signal (rBG PA).

23. A cell comprising the nucleic acid molecule of any one of the preceding claims.

24. The cell of claim 23 further comprising the enzyme capable of performing targeted genomic integration of the nucleic acid molecule, optionally wherein the enzyme capable of performing targeted genomic integration of the nucleic acid molecule is encoded by an RNA.

25. The cell of claim 23 or claim 24, wherein the cell is an immune cell, optionally wherein the cell is an immune cell selected from the group consisting of a T cell, a macrophage, and a natural killer (NK) cell.

26. The cell of any one of claims 23-25 further comprising an exogenous copy of CD47 and / or HLA-E, optionally wherein the exogenous copy of CD47 and / or HLA-E is integrated in the genome of the cell.

27. The cell of any one of claims 23-26, wherein the cell is administered to a subject, optionally to a mammalian subject, optionally to a human.

28. The cell of any one of claims 23-27, wherein the cell is a component of an allogeneic therapy.

29. A method of treating a hematological cancer in a subject, the method comprising administering the cell of any one of claims 23-28 to the subject, thereby treating the hematological cancer in the subject.

30. The method of claim 29, wherein the hematological cancer is selected from the group consisting of leukemia, lymphoma, and multiple myeloma.

31. A method of making a chimeric antigen receptor (CAR)-immune cell, the method comprising: obtaining an immune cell from a sample obtained from a subject, the sample comprising immune cells or immune cell progenitors; and transfecting the immune cell with a first nucleic acid molecule of any one of claims 1-22 and a second nucleic acid molecule encoding the enzyme capable of performing targeted genomic integration of the first nucleic acid molecule, optionally wherein the second nucleic acid molecule is RNA, to thereby create a transfected CAR-immune cell, wherein the immune cell is a T cell, a macrophage, or a natural killer (NK) cell.

32. A method of making a chimeric antigen receptor (CAR)-immune cell, the method comprising: administering to a subject a first nucleic acid molecule of any one of claims 1-22 and a second nucleic acid molecule encoding the enzyme capable of performing targeted genomic integration of the first nucleic acid molecule, optionally wherein the second nucleic acid molecule is RNA, wherein said administering uses lipid nanoparticles (LNPs) capable of directing the first and second nucleic acid molecules to an immune cell, to thereby create a CAR-immune cell, wherein the immune cell is a T cell, a macrophage, or a natural killer (NK) cell.

33. The method of claim 31 or 32, wherein the immune cell is a T cell.

34. The method of any one of claims 31-33, wherein the transfected CAR-immune cell is a CAR- T cell, optionally an autologous CAR-T cell or an allogeneic CAR-T cell.

35. The method of claim 31 or claim 32, wherein the immune cell is a macrophage.

36. The method of any one of claims 31, 32 or 35, wherein the transfected CAR-immune cell is a CAR-M cell, optionally an autologous CAR-M cell or an allogeneic CAR-M cell.

37. The method of claim 31 or claim 32, wherein the immune cell is a natural killer (NK) cell.

38. The method of any one of claims 31, 32 or 37, wherein the transfected CAR-immune cell is a CAR-NK cell, optionally an autologous CAR-NK cell or an allogeneic CAR-NK cell.

39. A nucleic acid of any one of SEQ ID NOs: 1-5, 13-65 and 76-80.

40. A cell comprising the nucleic acid of claim 39.

41. An amino acid sequence of any one of SEQ ID NOs: 6-11.

42. A cell comprising the amino acid sequence of claim 41.

43. A cell comprising a genomic integrant of the nucleic acid of any one of claims 1-22, wherein the genomic integrant is prepared by a process comprising contacting the cell with a Myotis lucifugus mobile element enzyme (MLT).

44. The cell of claim 43, wherein the genomic integrant is located at least 1 kb away from any promoter start site within the genome of the cell.

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