Nucleic acid-based delivery systems and uses thereof

Abstract

Compositions and methods are provided that include synthetic delivery systems that utilize polynucleotides. The polynucleotides disclosed herein can, for example, facilitate the binding, uptake, nuclear transport, and / or genomic integration of components of the synthetic delivery system, such as cargo. The compositions include a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system includes a polynucleotide that includes a recognition sequence that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

Description

[Technical Field]

[0001] cross reference This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 369,913, filed July 29, 2022, and U.S. Provisional Patent Application No. 63 / 506,800, filed June 7, 2023, each of which is incorporated herein by reference in its entirety. [Background technology]

[0002] background Transposable elements (TEs or transposons) are DNA sequences that can change location within the genome. The relationship of transposable elements to disease initiation is poorly understood.

[0003] Tumor-derived cell-free DNA may contain genetic alterations relevant to tumorigenesis, suggesting that circulating tumor-derived DNA (ctDNA) serves as a vehicle for genetic exchange between tumor cells. It has been suggested that ctDNA may transmit oncogenic mutations to reshape the tumor microenvironment. However, the underlying processes and mechanisms remain poorly understood. Summary of the Invention [Means for solving the problem]

[0004] Abstract Disclosed herein, in some embodiments, is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to any one of SEQ ID NOs: 73-75.

[0005] Disclosed herein, in some embodiments, is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 73.

[0006] Disclosed herein, in some embodiments, is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13 or 14.

[0007] Disclosed herein in some embodiments is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and the recognition sequence comprises (a) a nucleotide sequence having at least 90% sequence identity to at least 60 contiguous nucleotides of SEQ ID NO:33, or (b) a nucleotide sequence having at least 91% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:33.

[0008] Disclosed herein, in some embodiments, is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, wherein the recognition sequence comprises an AluSp, AluSx, AluSg2, ERV2 superfamily, or ERV3 superfamily transposable element, or a functional fragment thereof, that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell.

[0009] Disclosed herein, in some embodiments, is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and the polynucleotide comprises a nucleotide sequence having at least 86% sequence identity to any one of SEQ ID NOs: 7, 16, 25, 29, and 31.

[0010] In some embodiments, disclosed herein is a method for delivering cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by the target cell, and wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13 or 14.

[0011] Disclosed herein in some embodiments is a method of delivering cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by the target cell, and wherein the recognition sequence comprises (a) a nucleotide sequence having at least 90% sequence identity to at least 60 contiguous nucleotides of SEQ ID NO:33, or (b) a nucleotide sequence having at least 91% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:33.

[0012] Disclosed herein in some embodiments is a method of delivering cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, and the recognition sequence comprises an AluSp, AluSx, AluSg2, ERV2 superfamily, or ERV3 superfamily transposable element, or a functional fragment thereof, that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by the target cell.

[0013] Disclosed herein in some embodiments is a method for delivering cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by the target cell, and wherein the polynucleotide comprises a nucleotide sequence having at least 86% sequence identity to any one of SEQ ID NOs: 7, 16, 25, 29, and 31.

[0014] Disclosed herein in some embodiments is a method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13 or 14.

[0015] Disclosed herein in some embodiments is a method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and wherein the recognition sequence comprises (a) a nucleotide sequence having at least 90% sequence identity to at least 60 contiguous nucleotides of SEQ ID NO:33, or (b) a nucleotide sequence having at least 91% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:33.

[0016] Disclosed herein in some embodiments is a method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, and the recognition sequence comprises an AluSp, AluSx, AluSg2, ERV2 superfamily, or ERV3 superfamily transposable element, or a functional fragment thereof, that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell.

[0017] Disclosed herein in some embodiments is a method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and wherein the polynucleotide comprises a nucleotide sequence having at least 86% sequence identity to any one of SEQ ID NOs: 7, 16, 25, 29 and 31.

[0018] In some aspects, disclosed herein is a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide, and the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 10 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0019] In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 15 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 20 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 30 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 50 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 75 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 100 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 10 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 15 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 20 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 30 contiguous nucleotides of any one of SEQ ID NOs: 1-78.In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 50 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 75 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 100 contiguous nucleotides of any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide comprises the nucleotide sequence of any one of SEQ ID NOs: 1-78.

[0020] In some embodiments, the disclosure provides a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide, and the polynucleotide is selected from the group consisting of AluSp, MER11C, AluY, 2L2a, ALluY, ALR / alpha, ALU, AluJb, AluS1, AluSc8, AluSg, ALUSg2, AluSq, AluSq2, ALUSx, AluSx, AluSx1, AluSz, AluSz6, AluYc3, ASLUSq2, ERVK / LTR, ERVL, ERVL-MaLR, FLAM_ C and AluY, HERV17-int, HERV9N-int, L1M1, L1MB3, L1ME4b, L1MEg, L1P1, L1P3, L1PA10, L1PA15, L1PA7, L1PB4, L2a, L2a / LTR40b / MLT1J2, LINE / L1 , LINE / L2, LTR, LTR / ERV1, LTR / ERVL, LTR / ERVL-MaLR, LTR / Gypsy, LTR41C, LTR81B, Mam_R4, Mamr4, Many, MER11B, MER41E, MIR, MIRB, MIRc, MIRc- A pharmaceutical composition is provided that includes a transposable element that is part L2, MLT1J2, MLT2B1, MLT1J2, MLT2B4, parAluSp-FULLMTL1J2, REP522, satellite / centr, SINE / Alu, SINE / MIR, THE1A, THE1B, THE1C, or Tigger3a transposable element.

[0021] In some embodiments, the disclosure provides a pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide that directs migration of the synthetic delivery system to a target cell, wherein the polynucleotide comprises a nucleic acid sequence identified in circulating tumor DNA (ctDNA) from a cell type substantially similar to the target cell, and wherein the nucleic acid sequence comprises a short interspersed nucleotide sequence (SINE), a long interspersed nucleotide sequence (LINE), an ERVL, or an ERVK transposable element.

[0022] In some embodiments, the polynucleotide comprises an AluSp transposable element. In some embodiments, the polynucleotide comprises a MER11C transposable element. In some embodiments, the polynucleotide comprises an AluSx transposable element. In some embodiments, the polynucleotide comprises an MLT1J transposable element. In some embodiments, the polynucleotide comprises an AluSg2 transposable element. In some embodiments, the polynucleotide comprises a THE1A transposable element. In some embodiments, the polynucleotide comprises an AluJb transposable element. In some embodiments, the polynucleotide comprises an MLT2B4 transposable element. In some embodiments, the polynucleotide comprises an L2a transposable element. In some embodiments, the polynucleotide comprises an MLT1J2 transposable element. In some embodiments, the polynucleotide comprises an AluSq transposable element. In some embodiments, the polynucleotide comprises an L1MB3 transposable element. In some embodiments, the polynucleotide comprises a THE1C transposable element. In some embodiments, the polynucleotide comprises an AluY transposable element. In some embodiments, the polynucleotide is double-stranded DNA. In some embodiments, the synthetic delivery system further comprises a cargo. In some embodiments, the cargo comprises a nucleic acid cargo. In some embodiments, the nucleic acid cargo is attached to the 3' end of the polynucleotide. In some embodiments, the nucleic acid cargo is attached to the 5' end of the polynucleotide. In some embodiments, the synthetic delivery system comprises a promoter. In some embodiments, the nucleic acid cargo encodes a tumor suppressor protein. In some embodiments, the cargo comprises a cytotoxic cargo. In some embodiments, the cargo comprises a therapeutic cargo. In some embodiments, the synthetic delivery system does not utilize a viral vector, a nanoparticle, a lipid nanoparticle, a liposome, an exosome, a dendrimer, a gene gun, or electroporation.

[0023] In some embodiments, the present disclosure provides a method of delivering a cargo to a target cell, comprising contacting the target cell with the pharmaceutical composition or synthetic delivery system of any one of the preceding embodiments.

[0024] In some embodiments, the cargo is delivered to the nucleus of the target cell. In some embodiments, the synthetic delivery system further comprises a transposon integration signal. In some embodiments, the cargo comprises a nucleic acid cargo. In some embodiments, the nucleic acid cargo is integrated into the genome of the target cell. In some embodiments, the nucleic acid cargo is integrated into the genome of the target cell at a location identified in Table 2, Table 4, Table 7, or Table 8. In some embodiments, the nucleic acid cargo is integrated into the genome of the target cell at Chr2:32916224-32916626. In some embodiments, the nucleic acid cargo is integrated into the genome of the target cell at Chl6:32628381-32629000. In some embodiments, the target cell is a leukocyte. In some embodiments, the target cell is a plasma cell. In some embodiments, the target cell is a cancer cell. In some embodiments, the target cell is a multiple myeloma cell. In some embodiments, the target cell is a pancreatic cell. In some embodiments, the target cell is a pancreatic cancer cell. In some embodiments, the target cell is a gastrointestinal cell, hi some embodiments, the target cell is a colorectal cancer cell.

[0025] In some embodiments, the present disclosure provides a method of treating a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of the preceding embodiments. Incorporation by Reference

[0026] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference herein. [Brief explanation of the drawings]

[0027] [Figure 1-1]Figure 1A shows the acquisition of nucleotide variants in cells co-cultured with ctDNA. Comparative SNV analysis was performed between the cell genome, ctDNA, and ctDNA-cell co-culture. Venn diagrams display exclusive and shared SNVs between each experimental condition. Outlined areas highlight commonly shared SNVs between ctDNA and ctDNA / cell co-culture conditions.

[0028] [Figure 1-2] Figure 1B shows a stacked bar graph demonstrating the changes in allele depth of variant (light gray) and reference (dark gray) alleles in ctDNA, cell line genomes, and co-culture conditions. Cells under co-culture conditions have greater depth in variant alleles in some locations compared to the control cell genome.

[0029] [Figure 2-1] Figure 2A shows an image of the index IVG variant calls and their allele frequencies in the pancreatic cancer experimental condition. The horizontal bars represent the substituted nucleotides indicated below, and the light gray represents the reference nucleotide.

[0030] Figure 2B shows an image of the index IVG variant calls and their allele frequencies in the multiple myeloma experimental condition. The horizontal bars represent the substituted nucleotides indicated below, and the light gray represents the reference nucleotide.

[0031] [Figure 2-2] Figure 2C shows an image of the index IVG variant calls and their allele frequencies in the multiple myeloma experimental condition. The horizontal bars represent the substituted nucleotides indicated below, and the light gray represents the reference nucleotide.

[0032] [Figure 2-3] Figure 2D shows an image of the index IVG variant calls and their allele frequencies in the pancreatic cancer experimental condition. The horizontal bars represent the substituted nucleotides indicated below, and the light gray represents the reference nucleotide.

[0033] [Figure 3-1] Figure 3A shows BLAST alignment images demonstrating the transition points of insertions between cellular genome contigs (dark boxes) and ctDNA contigs (light boxes) for pancreatic cancer and multiple myeloma ctDNA. Results were obtained after comparing contigs carrying insertions in co-culture conditions with the reference cellular genome.

[0034] Figure 3A shows a BLAST alignment image demonstrating the transition points of insertions between the cellular genome contigs (dark boxes) and the ctDNA contigs (light boxes) for multiple myeloma ctDNA. Results were obtained after comparing the contigs carrying the insertions in the co-culture condition with the reference cellular genome.

[0035] [Figure 3-2] Figure 3A shows a BLAST alignment image demonstrating the transition points of insertions between the cellular genome contigs (dark boxes) and the ctDNA contigs (light boxes) for pancreatic cancer ctDNA. Results were obtained after comparing the contigs carrying insertions in co-culture conditions with the reference cellular genome.

[0036] [Figure 4] Figure 4 shows an overview of the distribution of transposon-containing contigs and the proportion of transposons observed at the 5' or 3' end in inserted versus non-inserted ctDNA fragments.

[0037] [Figure 5] Figure 5A shows illustrative common retrotransposon subfamilies and classes located at insertion points for pancreatic cancer ctDNA.

[0038] Figure 5B shows illustrative common retrotransposon subfamilies and classes located at insertion points for multiple myeloma ctDNA.

[0039] [Figure 6] FIG. 6A shows the expression levels of selected transposable elements in tumor samples.

[0040] FIG. 6B shows the expression levels of selected transposable elements in tumor samples.

[0041] [Figure 7] FIG. 7 shows the effect of reverse transcriptase inhibitors or integrase inhibitors on ctDNA chromatid integration.

[0042] [Figure 8] FIG. 8 shows an agarose gel of the transposon PCR products after incubation of the transposon construct in complete medium for 4 hours followed by PCR.

[0043] [Figure 9-1] Figure 9A shows the time course of Cy5-AluSp and Cy5-control sequence treated MM1s cells (1 μg / mL). CY5(+) cells were detected by flow cytometry.

[0044] Figure 9B shows the results of a titration experiment of Cy5-AluSp and Cy5-control sequence-treated MM1s cells. Prior to flow cytometry, half of the samples were treated with trypsin to determine the amount of internalized DNA.

[0045] [Figure 9-2] FIG. 9C shows the uptake and internalization of different multiple myeloma retrotransposons and controls by MM1s cells after 4 hours of culture, as assessed by flow cytometry.

[0046] FIG. 9D shows the uptake and internalization of different multiple myeloma retrotransposons and controls by U266 cells after 4 hours of culture, as assessed by flow cytometry.

[0047] [Figure 10]FIG. 10 provides illustrative flow cytometry scatter plots displaying cellular capture of AluSp, MER11, and control transposon sequences in plasma cells (CD138+) and non-plasma cells (CD138−) derived from the bone marrow of a multiple myeloma patient.

[0048] [Figure 11] Figure 11 shows the percentage of cells positive for fluorescently labeled AluSp, MER11, control sequences, or PC-specific transposon sequences after 14 hours of incubation with bone marrow from multiple myeloma patients or normal bone marrow. Data are shown separately for plasma cells (CD138+) and non-plasma cells (CD138-).

[0049] [Figure 12-1] Figure 12A illustrates the effect of 5' or 3' deletions on the internalization of Cy5-labeled AluSp by MM1s cells. Images were captured after culturing cells with retrotransposons for 8 hours.

[0050] Figure 12B provides a gel image showing the AluSp deletion.

[0051] [Figure 12-2] FIG. 12C is a graphic representation of adenine (A)-thymine (T) and guanine (G)-cytosine (C) enriched regions, or both, identified by multiple sequence alignment of AluSp and other MM-specific transposons.

[0052] [Figure 13] Figure 13 provides microscopy images of MM1s cells cultured with AluSp-CMV-mCherry, CMV-mCherry linear vector, or cells transfected with CMV-mCherry circular vector. Images were captured 24 hours after co-culture with DNA.

[0053] [Figure 14]FIG. 14A is a bar graph displaying a summary of the number of mCherry genomic insertions identified in MM1s cells expressing high levels of mCherry, intermediate levels of mCherry, or no mCherry (n=5 cells per group).

[0054] Figure 14B is an agarose gel showing PCR bands indicative of mCherry integration for chromatin extracted from cells treated with the AluSp-CMV-mCherry cassette or the control-CMV-mCherry cassette. *Nonspecific band.

[0055] FIG. 14C displays the confidence of detection of the identified insertion versus the number of samples in which the specific site of insertion was detected.

[0056] [Figure 15] Figure 15 shows cell viability of three different cell lines co-cultured with TE-HSV-Tk-GFP for 24 hours before adding ganciclovir (GCV). Apoptosis was measured 96 hours after GCV addition. MM: multiple myeloma, CC: colon cancer, and PC: pancreatic cancer. TE-CMV-GFP: transposon element linked to CMV-GFP. HSV-TK: herpes simplex virus thymidine kinase. Error bars in box plots indicate standard deviations of triplicate experiments.

[0057] [Figure 16] Figure 16 shows the results of a cell viability assay measuring sensitivity to gemcitabine in pancreatic cancer cell lines (MIA and ASPC-1) cultured with plasma from patients resistant to gemcitabine, similar plasma pretreated with DNase I, or plasma from control non-cancer patients. For the corresponding DNase I-treated samples, plasma was treated with DNase I for 10 minutes. GR: gemcitabine resistant.

[0058] [Figure 17]Figure 17 shows a comparison of the cell viability response to bortezomib in OPM1 cells cultured with plasma from a patient who failed to respond to bortezomib treatment, a combination of ctDNA from the same patient who was resistant to bortezomib and control plasma, and control plasma alone (non-cancer patient) (top left); the viability response of RPMI to control plasma or control plasma spiked with ctDNA from a patient who achieved a complete response to bortezomib (center); and cell viability assessment after bortezomib treatment of MM1s cells cultured with plasma from a bortezomib-resistant patient (BR#2) alone, or with BR#2 alone after treatment with DNase I or co-culture with ctDNA from a different bortezomib-resistant patient (BR#1) (right). BR: bortezomib-resistant; BS: bortezomib-sensitive. Error bars indicate standard deviation of triplicate experiments.

[0059] [Figure 18-1] Figure 18A provides a bar graph comparing the results of cell capture of synthesized CY5-labeled retrotransposon sequences derived from multiple myeloma (MM) ctDNA with controls. Data are grouped by cell line. In each case, the bars presented are, from left to right: control followed by synthesized MM-derived DNA (MM01, MM02, MM03, MM04, and ZIP2). The horizontal line defines the control cutoff.

[0060] [Figure 18-2] Figure 18B provides a bar graph comparing the results of cell capture of synthesized CY5-labeled retrotransposon sequences derived from pancreatic ductal carcinoma (PDAC) ctDNA with controls. Data are grouped by cell line. In each case, the bars presented are, from left to right: control followed by synthesized PDAC-derived DNA (PC01, PC02, PC03, PC21, PC22, PC23, PC24). The horizontal line defines the control cutoff.

[0061] [Figure 19]Figure 19 provides fluorescence microscopy images showing the uptake of fluorescently labeled synthetic DNA derived from multiple myeloma ctDNA (MM2(1-290), MM2(89-290), MM2(177-290), and MM2(1-201) and MM2(1-113)) or control sequences.

[0062] [Figure 20] Figure 20 provides fluorescence microscopy images assessing the uptake of fluorescently labeled synthetic DNA derived from multiple myeloma ctDNA (MM2(1-290), MM2(89-290), MM2(177-290), and MM2(1-201) and MM2(1-113)) by multiple myeloma / B cell lines (MM1S and RPMI) and other cell types (HFF - fibroblast, DU145 - prostate cancer, MCF10A breast).

[0063] [Figure 21] FIG. 21 provides microscopy images showing the uptake of 5′ fluorescently labeled synthetic DNA fragments derived from multiple myeloma ctDNA by MM cells (JK6L) or non-MM cells. DETAILED DESCRIPTION OF THE INVENTION

[0064] Detailed Description Compositions and methods are provided that include synthetic delivery systems that utilize the polynucleotides disclosed herein. The polynucleotides disclosed herein can, for example, facilitate binding, uptake, nuclear transport, and / or genomic integration of components of the synthetic delivery system, such as cargo.

[0065] The synthetic delivery systems disclosed herein can utilize polynucleotides that facilitate cell-specific horizontal gene transfer (HGT) between cells (e.g., human cancer cells). Gene transfer between cells can play physiological and pathological roles in certain organisms. For example, HGT has been characterized in prokaryotes, according to HGT can provide survival and evolutionary advantages against environmental stressors. Evidence for HGT in mammals and humans is much more limited, as is our understanding of the underlying mechanisms involved.

[0066] The present invention identifies tissue-specific retrotransposons that can mediate the process by which circulating tumor DNA (ctDNA) targets and spreads genetic material to cells that resemble the cells of origin of ctDNA.The compositions and methods provided herein include synthetic delivery systems that utilize components, such as polynucleotides, to facilitate cargo binding, uptake, nuclear transport and / or genome integration.The compositions and methods can be used to deliver cargo to specific cell types and cells, including cancer cells. I. Synthetic delivery system

[0067] For example, synthetic delivery systems are disclosed herein that can provide cell-, tissue-, and / or organ-specific targeting, delivery, binding, uptake, nuclear localization, and / or genomic integration of cargo. The compositions and methods provided herein include synthetic delivery systems that utilize components of the synthetic delivery system, such as polynucleotides, to facilitate cargo binding, uptake, nuclear transport, and / or genomic integration. The compositions and methods can be used to deliver cargo to specific cell types and cells, including cancer cells.

[0068] The synthetic delivery system disclosed herein can comprise a polynucleotide. The synthetic delivery system can comprise, for example, a polynucleotide and a cargo. The polynucleotide can be or comprise double-stranded DNA (dsDNA). In some embodiments, the polynucleotide comprises single-stranded DNA. In some embodiments, the polynucleotide comprises RNA.

[0069] The polynucleotides of the present disclosure, their sequences or portions thereof (e.g., recognition and / or integration sequences) can be derived from or from nucleic acid sequences of a biological sample (e.g., a blood, plasma, or tissue sample of a subject, such as a human). The polynucleotides of the present disclosure, their sequences or portions thereof (e.g., recognition and / or integration sequences) can be derived from or from cancerous cells or tissues. The polynucleotides can be recombinant or synthetic, e.g., not isolated from a biological sample.

[0070] The polynucleotide or a portion thereof (e.g., recognition and / or integration sequence) in the synthetic delivery system can be derived from or derived from, for example, circulating tumor DNA (ctDNA) from a human. The ctDNA can comprise double-stranded DNA (dsDNA). In some embodiments, the ctDNA comprises single-stranded DNA. Such ctDNA or a fragment thereof can comprise a nucleic acid sequence that can target a specific cell or cell population and induce binding, uptake, nuclear localization, and / or genomic integration of the polynucleotide or cargo disclosed herein.

[0071] The polynucleotide in the synthetic delivery system that delivers a cargo to a target cell can comprise a nucleic acid sequence derived from or derived from ctDNA. The ctDNA can originate from the same cell type or tissue type as the target cell. The polynucleotide used to deliver a cargo to a target cell can comprise a nucleic acid sequence derived from or derived from ctDNA of substantially the same cell type or substantially the same tissue type as the target cell. The polynucleotide used to deliver a cargo to a target cell can comprise a nucleic acid sequence derived from or derived from ctDNA of substantially the same cell type or substantially the same tissue type as the target cell. For example, ctDNA derived from multiple myeloma or a component derived therefrom (e.g., a subsequence or recognition sequence) can be used to generate a synthetic delivery system for delivery, binding, uptake, nuclear localization, and / or genomic integration of a cargo into multiple myeloma target cells or related target cells or target cell types. Pancreatic cancer-derived ctDNA or components derived therefrom (e.g., subsequences or recognition sequences) can be used to generate synthetic delivery systems for delivery, binding, uptake, nuclear localization, and / or genomic integration of cargo into pancreatic cancer target cells or related target cells or cell types. Colorectal cancer-derived ctDNA or components derived therefrom (e.g., subsequences or recognition sequences) can be used to generate synthetic delivery systems for delivery, binding, uptake, nuclear localization, and / or genomic integration of cargo into colorectal cancer target cells or related target cells or cell types. Lung cancer-derived ctDNA or components derived therefrom (e.g., subsequences or recognition sequences) can be used to generate synthetic delivery systems for delivery, binding, uptake, nuclear localization, and / or genomic integration of cargo into lung cancer target cells or related target cells or cell types.

[0072] The synthetic delivery system, polynucleotide, nucleic acid sequence, recognition sequence, integration sequence, or cargo can comprise natural or non-natural nucleotide analogs or bases, or combinations thereof. The nucleotide analogs can contain modification(s) in one or more of the ribose moiety, phosphate moiety, nucleoside moiety, or combinations thereof. Nucleotide analogs or bases can include 2'-O-methyl, 2'-O-methoxyethyl (2'-O-MOE), 2'-O-aminopropyl, 2'-deoxy, T-deoxy-2'-fluoro, 2'-O-aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-O-DMAOE), 2'-O-dimethylaminopropyl (2'-O-DMAP), T-dimethylaminoethyloxyethyl (2'-O-DMAEOE), or 2'-ON-methylacetamide (2'-O-NMA) modifications, locked nucleic acids (LNA), ethylene nucleic acids (ENA), peptide nucleic acids (PNA), 1',5'-anhydrohexitol nucleic acids (HNA), morpholinos, methylphosphonate nucleotides, thiolphosphonate nucleotides, or 2'-fluoro N3-P5'-phosphoramidites. Polynucleotides can include one or more abasic sites. The abasic site can be functionalized with a detectable moiety.

[0073] The synthetic delivery system, polynucleotide, first nucleic acid sequence, or second nucleic acid sequence can include a recognition sequence that can provide cell-specific targeting and / or uptake of the polynucleotide, synthetic delivery system, or cargo. In some embodiments, the recognition sequence binds to a receptor that mediates or is associated with the uptake of ctDNA and / or the synthetic delivery system. In some embodiments, the recognition sequence of the ctDNA, synthetic delivery system, or polynucleotide specifically or preferentially binds to a receptor associated with the uptake of ctDNA and / or the synthetic delivery system.

[0074] In some embodiments, the recognition sequence facilitates preferential binding of the synthetic delivery system, polynucleotide, or cargo to the target cell compared to a control cell. In some embodiments, the recognition sequence facilitates preferential uptake of the ctDNA, synthetic delivery system, polynucleotide, or cargo by the target cell compared to a control cell. In some embodiments, the recognition sequence facilitates preferential binding, uptake, nuclear localization, and / or genomic integration of the ctDNA, synthetic delivery system, polynucleotide, or cargo by the target cell compared to a control cell. The control cell can be, for example, a cell of a different cell type (e.g., a cell type disclosed herein). In some embodiments, the target cell is a cancer cell and the control cell is a non-cancer cell. In some embodiments, the target cell is a cancer cell of a first cell type and the control cell is a cancer cell of a second cell type. In some embodiments, the target cell is a cancer cell of a first cell type and the control cell is a non-cancer cell of a second cell type.

[0075] The recognition sequence may be or comprise a functional fragment and / or variant of the sequence disclosed herein. For example, the recognition sequence may comprise a subset of nucleotides derived from the nucleotide sequence disclosed herein that facilitates preferential binding to target cells compared to control cells, preferential uptake by target cells compared to control cells, preferential nuclear localization compared to control cells, and / or preferential integration into the genome of target cells compared to control cells. In some embodiments, the recognition sequence comprises a functional fragment and / or variant of the sequence disclosed herein that retains at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the target cell binding, uptake, nuclear localization, and / or genome integration activity of the nucleotide sequence from which the fragment and / or variant is derived.

[0076] For example, in some embodiments, the synthetic delivery systems, polynucleotides, nucleic acid sequences, first nucleic acid sequences, second nucleic acid sequences, recognition sequences, or integration sequences disclosed herein comprise a subset of nucleotides from SEQ ID NO:68 or SEQ ID NO:78 that facilitate, e.g., preferential binding to target cells compared to control cells, preferential uptake by target cells compared to control cells, preferential nuclear localization compared to control cells, and / or preferential integration into the genome of target cells compared to control cells, e.g., SEQ ID NO:68 or SEQ ID NO:78. [ka] [ka] In some embodiments, the recognition sequence comprises a functional fragment and / or variant of SEQ ID NO:68 or SEQ ID NO:78 that retains at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the target cell binding, uptake, nuclear localization, and / or genome integration activity of SEQ ID NO:68 or SEQ ID NO:78.

[0077] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence does not contain a nucleotide sequence having at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, at least 97%, or at least 99% sequence identity to nucleotides 1-87 of SEQ ID NO: 68 or nucleotides 1-87 of SEQ ID NO: 78. In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence does not contain the nucleotide sequence of nucleotides 1-87 of SEQ ID NO: 68 or nucleotides 1-87 of SEQ ID NO: 78. In some embodiments, the synthetic delivery system or polynucleotide comprises the nucleotide sequence of nucleotides 1-87 of SEQ ID NO:68, nucleotides 1-87 of SEQ ID NO:78, or a sequence having at least minimal sequence identity thereto, but said sequence is not part of a recognition sequence, e.g., is not required for preferential binding to, uptake by, or nuclear localization in, a target cell.

[0078] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence does not contain a nucleotide sequence having at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 71. In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence does not contain the nucleotide sequence of SEQ ID NO: 71. In some embodiments, the synthetic delivery system or polynucleotide comprises the nucleotide sequence of SEQ ID NO: 71, or a sequence having at least minimal sequence identity thereto, but said sequence is not part of the recognition sequence, e.g., is not required for preferential binding to, uptake by, or nuclear localization in, a target cell.

[0079] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises at most 10%, at most 20%, at most 30%, at most 40%, at most 50%, at most 60%, at most 70%, at most 80% or at most 90% sequence identity to SEQ ID NO:68 or SEQ ID NO:78.

[0080] The recognition sequence may be a subset of contiguous nucleotides from the sequences disclosed herein, e.g., at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, or at least about 200, or at least about 210, or at least about 220, or at least about 230, or at least about 240, or at least about 250, or at least about 260, or at least about 270, or at least about 280, or at least about 290, or at least about 300, or at least about 310, or at least about 320, or at least about 330, or at least about 340, or at least about 350, or at least about 360, or at least about 370, or at least about 380, or at least about 390, or at least about 400, or at least about 410, or at least about 420, or at least about 430, or at least about 440, or at least about 450, or at least about 460, or at least about 470, or at least about 480, or at least about 490, or at least about 500, or at least about 510, or at least about 520, or at least about 530, or at least about 540, or at least about 550, or at least about 560, or at least , at least about 200, at most about 10, at most about 20, at most about 30, at most about 40, at most about 50, at most about 60, at most about 70, at most about 80, at most about 90, at most about 100, at most about 110, at most about 120, at most about 130, at most about 140, at most about 150, at most about 160, at most about 170, at most about 180, at most about 190, at most about 200, at most about 250, at most about 300, at most about 350, at most about 400, at most about 450, at most about 500, about 10, about 15, about 20, about 2 5, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 5-10, about 5-2 0, about 5-30, about 5-40, about 5-50, about 5-60, about 5-70, about 5-80, about 5-90, about 5-100, about 5-150, about 5-200, about 5-300, about 5-400, about 5-500, about 10-20, about 10-30, about 10-40, about 10-50, about 10-60, about 10- 70, approximately 10-80, approximately 10-90, approximately 10-100, approximately 10-150, approximately 10-200, approximately 10-300, approximately 10-400, approximately 10-500, approximately 15-20, approximately 15-30, approximately 15-40, approximately 15-50, approximately 15-60, approximately 15-70, approximately 15-80, approximately 15-90, approximately 15- 100, approximately 15-150, approximately 15-200, approximately 15-300, approximately 15-400, approximately 15-500, approximately 20-30, approximately 20-40, approximately 20-50, approximately 20-60, approximately 20-70, approximately 20-80, approximately 20-90, approximately 20-100, approximately 20-150, approximately 20-200, approximately 20-300,Approximately 20-400, approximately 20-500, approximately 30-40, approximately 30-50, approximately 30-60, approximately 30-70, approximately 30-80, approximately 30-90, approximately 30-100, approximately 30-150, approximately 30-200, approximately 30-300, approximately 30-400, approximately 30-500, approximately 40-50, approximately 40-60, approximately 40-70, approximately 40-80, approximately 40-90, approximately 40-100, approximately 40-150, approximately 40-200, approximately 40-300, approximately 40-400, approximately 40-500, approximately 50 It can comprise, consist essentially of, or consist of about 60, about 50 to 70, about 50 to 80, about 50 to 90, about 50 to 100, about 50 to 150, about 50 to 200, about 50 to 300, about 50 to 400, about 50 to 500, about 100 to 150, about 100 to 200, about 100 to 300, about 100 to 400, about 100 to 500, about 200 to 300, about 200 to 400, or about 200 to 500 consecutive nucleotides.

[0081] A recognition sequence can comprise, consist essentially of, or consist of a nucleotide sequence having at least a minimum percentage sequence identity to a subset of contiguous nucleotides from the sequences disclosed herein.

[0082] In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0083] In some embodiments, the recognition sequence comprises at least 80% sequence identity for at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 contiguous nucleotides from any one of SEQ ID NOs: 1-78.

[0084] In some embodiments, the recognition sequence comprises at least 85% sequence identity for at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0085] In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0086] In some embodiments, the recognition sequence comprises at least 95% sequence identity over at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 contiguous nucleotides from any one of SEQ ID NOs: 1-78.

[0087] In some embodiments, the recognition sequence comprises at least 98% sequence identity over at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0088] In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:73.

[0089] In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:74.

[0090] In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 125 contiguous nucleotides of SEQ ID NO: 75. In some embodiments, the recognition sequence comprises at least 70% sequence identity to at least about 150 contiguous nucleotides of SEQ ID NO: 75.

[0091] In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:73.

[0092] In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:74.

[0093] In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 125 contiguous nucleotides of SEQ ID NO: 75. In some embodiments, the recognition sequence comprises at least 80% sequence identity to at least about 150 contiguous nucleotides of SEQ ID NO: 75.

[0094] In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:73.

[0095] In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:74.

[0096] In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 125 contiguous nucleotides of SEQ ID NO: 75. In some embodiments, the recognition sequence comprises at least 90% sequence identity to at least about 150 contiguous nucleotides of SEQ ID NO: 75.

[0097] In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:73. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:73.

[0098] In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:74.

[0099] In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 30 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 40 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 50 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 60 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 70 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 80 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 90 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 100 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 125 contiguous nucleotides of SEQ ID NO: 75. In some embodiments, the recognition sequence comprises at least 95% sequence identity to at least about 150 contiguous nucleotides of SEQ ID NO: 75.

[0100] In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 30 contiguous nucleotides of SEQ ID NO: 73. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 40 contiguous nucleotides of SEQ ID NO: 73. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 50 contiguous nucleotides of SEQ ID NO: 73. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 60 contiguous nucleotides of SEQ ID NO: 73. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 70 contiguous nucleotides of SEQ ID NO: 73. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 80 contiguous nucleotides of SEQ ID NO: 73.

[0101] In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 30 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 40 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 50 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 60 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 70 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 80 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 90 contiguous nucleotides of SEQ ID NO:74. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 100 contiguous nucleotides of SEQ ID NO:74.

[0102] In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 30 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 40 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 50 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 60 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 70 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 80 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 90 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 100 contiguous nucleotides of SEQ ID NO:75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 125 contiguous nucleotides of SEQ ID NO: 75. In some embodiments, the recognition sequence comprises, consists essentially of, or consists of at least about 150 contiguous nucleotides of SEQ ID NO: 75.

[0103] In some embodiments, the recognition sequence is at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, or at least about 200 nucleotides in length.

[0104] In some embodiments, the recognition sequence is at most about 10, at most about 20, at most about 30, at most about 40, at most about 50, at most about 60, at most about 70, at most about 80, at most about 90, at most about 100, at most about 110, at most about 120, at most about 130, at most about 140, at most about 150, at most about 160, at most about 170, at most about 180, at most about 190, at most about 200, at most about 250, at most about 300, at most about 350, at most about 400, at most about 450, or at most about 500 nucleotides in length.

[0105] In some embodiments, the recognition sequence is about 10, about 15, about 20, about 25, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 250, about 300, about 350, about 400, about 450, or about 500 nucleotides in length.

[0106] In some embodiments, the recognition sequence is about 5 to 10, about 5 to 20, about 5 to 30, about 5 to 40, about 5 to 50, about 5 to 60, about 5 to 70, about 5 to 80, about 5 to 90, about 5 to 100, about 5 to 150, about 5 to 200, about 5 to 300, about 5 to 400, about 5 to 500, about 10 to 20, about 10 to 30, about 10 to 40, about 10 to 50, about 10 to 60, about 10 to 70, about 10 to 80, about 10 to 90, about 10 to 100, about 10 to 150, about 10 ...0, about 10 to 300, about 10 to 400, about 10 to 500, about 10 to 200, about 10 to 300, about 10 to 400, about 10 to 500, about 10 to 200, about 10 to 300, about 10 to 400, about 10 to 500 200, about 10-300, about 10-400, about 10-500, about 15-20, about 15-30, about 15-40, about 15-50, about 15-60, about 15-70, about 15-80, about 15-90, about 15-100, about 15-150, about 15-200, about 15-300, about 15-400, about 15-500, about 20-30, about 20-40, about 20-50, about 20-60, about 20-70, about 20-80, about 20-90, about 20-100 , about 20-150, about 20-200, about 20-300, about 20-400, about 20-500, about 30-40, about 30-50, about 30-60, about 30-70, about 30-80, about 30-90, about 30-100, about 30-150, about 30-200, about 30-300, about 30-400, about 30-500, about 40-50, about 40-60, about 40-70, about 40-80, about 40-90, about 40-100, about 40-150, about 40-20 The length is 0, about 40 to 300, about 40 to 400, about 40 to 500, about 50 to 60, about 50 to 70, about 50 to 80, about 50 to 90, about 50 to 100, about 50 to 150, about 50 to 200, about 50 to 300, about 50 to 400, about 50 to 500, about 100 to 150, about 100 to 200, about 100 to 300, about 100 to 400, about 100 to 500, about 200 to 300, about 200 to 400, or about 200 to 500 nucleotides.

[0107] In some embodiments, the recognition sequence of the synthetic delivery system or polynucleotide is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, at least 20-fold, at least 21-fold, at least 22-fold, at least 23-fold, at least 24-fold, at least 25-fold, at least 26-fold, at least 27-fold, at least 28-fold, at least 29-fold, at least 30-fold, at least 31-fold, at least 32-fold, at least 33-fold, at least 34-fold, at least 35-fold, at least 36-fold, at least 37-fold, at least 38-fold, at least 39-fold, at least 40-fold, at least 41-fold, at least 42-fold, at least 43-fold, at least 44-fold, at least 45-fold, at least 46-fold, at least 47-fold, at least 48-fold, at least 49-fold, at least 50-fold, at least 51-fold, at least 52-fold, at least 53-fold, at least 54-fold, at least 55-fold, at least 56-fold, at least 57-fold, at facilitates binding to a target cell (e.g., a population thereof) as disclosed herein that is 5-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 500-fold, at least 600-fold, at least 700-fold, at least 800-fold, at least 900-fold, at least 1000-fold, at least 1500-fold, at least 2000-fold, or at least 3000-fold greater.

[0108] In some embodiments, the recognition sequence of the synthetic delivery system or polynucleotide is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, at least 20-fold, at least 21-fold, at least 22-fold, at least 23-fold, at least 24-fold, at least 25-fold, at least 26-fold, at least 27-fold, at least 28-fold, at least 29-fold, at least 30-fold, at least 31-fold, at least 32-fold, at least 33-fold, at least 34-fold, at least 35-fold, at least 36-fold, at least 37-fold, at least 38-fold, at least 39-fold, at least 40-fold, at least 41-fold, at least 42-fold, at least 43-fold, at least 44-fold, at least 45-fold, at least 46-fold, at least 47-fold, at least 48-fold, at least 49-fold, at least 50-fold, at least 51-fold, at least 52-fold, at least 53-fold, at least 54-fold, at least 55-fold, at least 56-fold, at least 57-fold, at facilitates uptake of the synthetic delivery system, polynucleotide or cargo by target cells that is 5 fold, at least 20 fold, at least 30 fold, at least 40 fold, at least 50 fold, at least 60 fold, at least 70 fold, at least 80 fold, at least 90 fold, at least 100 fold, at least 150 fold, at least 200 fold, at least 250 fold, at least 300 fold, at least 350 fold, at least 400 fold, at least 500 fold, at least 600 fold, at least 700 fold, at least 800 fold, at least 900 fold, at least 1000 fold, at least 1500 fold, at least 2000 fold, or at least 3000 fold greater.

[0109] In some embodiments, the recognition sequence of the synthetic delivery system or polynucleotide is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, at least 20-fold, at least 21-fold, at least 22-fold, at least 23-fold, at least 24-fold, at least 25-fold, at least 26-fold, at least 27-fold, at least 28-fold, at least 29-fold, at least 30-fold, at least 31-fold, at least 32-fold, at least 33-fold, at least 34-fold, at least 35-fold, at least 36-fold, at least 37-fold, at least 38-fold, at least 39-fold, at least 40-fold, at least 41-fold, at least 42-fold, at least 43-fold, at least 44-fold, at least 45-fold, at least 46-fold, at least 47-fold, at least 48-fold, at least 49-fold, at least 50-fold, at least 51-fold, at least 52-fold, at least 53-fold, at least 54-fold, at least 55-fold, at least 56-fold, at least 57-fold, at fold, at least 20 fold, at least 30 fold, at least 40 fold, at least 50 fold, at least 60 fold, at least 70 fold, at least 80 fold, at least 90 fold, at least 100 fold, at least 150 fold, at least 200 fold, at least 250 fold, at least 300 fold, at least 350 fold, at least 400 fold, at least 500 fold, at least 600 fold, at least 700 fold, at least 800 fold, at least 900 fold, at least 1000 fold, at least 1500 fold, at least 2000 fold, or at least 3000 fold greater, facilitates nuclear localization of the synthetic delivery system, polynucleotide or cargo in the target cell.

[0110] In some embodiments, the recognition sequence of the synthetic delivery system or polynucleotide is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, at least 20-fold, at least 21-fold, at least 22-fold, at least 23-fold, at least 24-fold, at least 25-fold, at least 26-fold, at least 27-fold, at least 28-fold, at least 29-fold, at least 30-fold, at least 31-fold, at least 32-fold, at least 33-fold, at least 34-fold, at least 35-fold, at least 36-fold, at least 37-fold, at least 38-fold, at least 39-fold, at least 40-fold, at least 41-fold, at least 42-fold, at least 43-fold, at least 44-fold, at least 45-fold, at least 46-fold, at least 47-fold, at least 48-fold, at least 49-fold, at least 50-fold, at least 51-fold, at least 52-fold, at least 53-fold, at least 54-fold, at least 55-fold, at least 56-fold, at least 57-fold, at fold, at least 20 fold, at least 30 fold, at least 40 fold, at least 50 fold, at least 60 fold, at least 70 fold, at least 80 fold, at least 90 fold, at least 100 fold, at least 150 fold, at least 200 fold, at least 250 fold, at least 300 fold, at least 350 fold, at least 400 fold, at least 500 fold, at least 600 fold, at least 700 fold, at least 800 fold, at least 900 fold, at least 1000 fold, at least 1500 fold, at least 2000 fold, or at least 3000 fold greater facilitates genomic integration of the synthetic delivery system, polynucleotide or cargo by the target cell.

[0111] In some instances, a multiple myeloma (MM)-derived recognition sequence can target binding and / or uptake of a synthetic delivery system, polynucleotide and / or cargo by MM cells, but not by, for example, pancreatic cells. A pancreatic cancer (PC)-derived recognition sequence can target binding and / or uptake of a synthetic delivery system, polynucleotide and / or cargo by pancreatic cancer cells, but not by, for example, multiple myeloma cells.

[0112] The recognition sequence can be derived from ctDNA. For example, the recognition sequence can be a fragment of ctDNA identified by sequencing ctDNA from a subject, for example, from the plasma of a human subject with cancer. The recognition sequence can be derived from ctDNA originating from lymphoma, leukemia, lung cancer, colon cancer (e.g., colorectal cancer), brain cancer, multiple myeloma, pancreatic cancer, hematological cancer, solid cancer, or other cancer types disclosed herein.

[0113] The synthetic delivery system or polynucleotide can include any suitable number of recognition sequences, for example, 1, 2, 3, 4, or 5 recognition sequences. In some embodiments, the synthetic delivery system or polynucleotide includes one recognition sequence. In some embodiments, the synthetic delivery system or polynucleotide includes two recognition sequences, for example, a first recognition sequence in the 5' region of the polynucleotide and a second recognition sequence in the 3' region of the polynucleotide, optionally separated by a non-recognition sequence or component, such as a nucleic acid cargo.

[0114] In some embodiments, the synthetic delivery system or polynucleotide comprises at least two, at least three, at least four, at least five, or another suitable number of recognition sequences at either the 3' or 5' end of the polynucleotide, or a combination thereof. The compositions of the present disclosure can comprise multiple such polynucleotides, and each polynucleotide can target one or more cell type(s). Thus, the methods and compositions of the present disclosure can be used to target one or more different target cells, one or more different target cell population(s), and / or one or more different target tissue(s), for example, simultaneously.

[0115] The synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) can comprise a transposable element (TE) or a derivative thereof, e.g., a fragment having a functional attribute disclosed herein. The transposable element can comprise a transposon or a derivative thereof. The transposable element can comprise a retrotransposon or a derivative thereof. The transposable element or transposon can be of any suitable class, subclass, family, subfamily, type, or others, including those disclosed herein. The transposable element or transposon can be a variant or derivative of a transposable element of a given class, subclass, family, subfamily, or type, including those disclosed herein.

[0116] The synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) can comprise, for example, a transposable element or fragment thereof derived from or from a class I or class II transposon. In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) comprises a transposable element or fragment thereof derived from or from a short interspersed nucleotide element (SINE), a long interspersed nucleotide element (LINE), an ERVL, or an ERVK transposable element.

[0117] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) is selected from the group consisting of AluSp, MER11C, AluY, 2L2a, ALluY, ALR / alpha, ALU, AluJb, AluS1, AluSc8, AluSg, ALUSg2, AluSq, AluSq2, ALUSx, AluSx, AluSx1, AluSz, AluSz6, AluYc3, ASLUSq2, ERVK / LTR, ERVL, ERVL-MaLR, FLAM_C, AluY, FLAM_C and AluY, HERV17-int, HERV9N-int, L1M1, L1MB3, L1ME4b, L1MEg, L1P1, L1P3, L1PA10, L1PA15, L1PA7, L1PB4, L2a, L2a / LTR40b / MLT1J2, LINE / L1, L INE / L2, LTR, LTR / ERV1, LTR / ERVL, LTR / ERVL-MaLR, LTR / Gypsy, LTR41C, LTR81B, Mam_R4, Mamr4, Many, MER11B, MER41E, MIR, MIRB, MIRc, MIRc-part L2, MLT1J2, MLT2B1, MLT1J2, MLT2B4, parAluSp-FULLMTL1J2, REP522, satellite / center, SINE / Alu, SINE / MIR, THE1A, THE1B, THE1C or Tigger3a transposable element or a fragment thereof derived from or from a transposable element or a derivative thereof.

[0118] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., a recognition sequence, an integration sequence, a first nucleic acid sequence, or a second nucleic acid sequence) comprises a transposable element or fragment thereof that is derived from or from a non-LTR retrotransposon transposable element, such as a SINE, LINE, or composite transposable element.

[0119] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or a fragment thereof that is derived from or from a LINE-class transposable element.In some embodiments, the transposable element or fragment thereof is a LINE class transposable element of the L1 group, e.g., L1, L1HS, L1M1B_5, L1M1_5, L1M2A1_5, L1M2A_5, L1M2B_5, L1M2C_5, L1M2_5, L1M3A_5, L1M3B_5, L1M3C_5, L1M3DE_5, L1M3D_5, L1M4B, L1M6B_5end, L1M6_5end, L1M7_5end, L1MA1, L1MA10, L1MA2, L1MA3, L1MA4, L1MA4A, L1MA5, L1MA5A, L1MA6, L1MA7, L1MA8, L1MA9, L1MA9_5, L1MB1, L1MB2, L1 MB3, L1MB3_5, L1 MB4, L1MB4_5, L1 MB5, L1MB6_5, L1 MB7, L1 MB8, L1MC1, L1MC2, L1MC3, L1MC4, L1MC4B, L1MC4_5end, L1MC5, L1MCA_5, L1MCB_5, L1MCC_5, L1MD1, L1MD1_5, L1MD2, L1MD3, L1MDA_5, L1MDB_5, L1ME1, L1 ME2, L1ME3, L1ME3A, L1ME3C_3end, L1ME3D_3end, L1ME3E_3end, L1ME3F_3end, L1ME4, L1ME4A, L1ME5, L1ME5_3end, L1MEA_5, L1MEB_5, L1MEC_5, L1MED_5 , L1MEE_5, L1MEe_5end, L1MEf_5end, L1MEg_5end, L1ME_ORF2, L1P4a_5end, L1P4b_5end, L1P4c_5end, L1P4d_5end, L1P4e_5end, L1PA10, L1PA11, L1PA1 2, L1PA12_5, L1PA13, L1PA13_5, L1PA14, L1PA14_5, L1PA15, L1PA16, L1PA16_5, L1PA17_5, L1PA2, L1PA3, L1PA4, L1PA5, L1PA6, L1PA7, L1PA7_5, L1PA8, L1 It is or is derived from a PB1, L1 PB2, L1PB2c, L1 PB3, L1 PB4, L1PBA1_5, L1PBA_5, L1PBB_5, L1PREC1, L1PREC2, L1P_MA2, HAL1, HAL1B (HAL1b), HAL1M8, IN25, MER25 or X9_LINE transposable element.In some embodiments, the transposable element or fragment thereof is or is derived from a LINE class transposable element of the L2 group, e.g., an L2, L2A, L2B, L2C, L2D, X15_LINE, X24_LINE, UCON49, or UCON86 transposable element.

[0120] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or a fragment thereof derived from or from a SINE-class transposable element. In some embodiments, the transposable element or fragment thereof is or is derived from a SINE-class transposable element of the SINE1 / 7S group, e.g., an AluY-series, AluS-series, AluJ-series, or monomeric Alu-series transposable element.

[0121] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) is an AluY series transposable element, e.g., ALU, AluY, AluYa1, AluYa4, AluYa5, AluYa8, AluYb10, AluYb11, AluYb3a1, AluYb3a2, AluYb8, AluYb8a1, AluYb9, AluYb10, AluYb11, AluYb12, AluYb13, AluYb14, AluYb15, AluYb16, AluYb17, AluYb18, AluYb19, AluYb20, AluYb21, AluYb22, AluYb23, AluYb24, AluYb25, AluYb26, AluYb27, AluYb28, AluYb29, AluYb30, AluYb31, AluYb32, AluYb33, AluYb34, AluYb35, AluYb36, AluYb37, AluYb38, AluYb39, AluYb40, AluYb41, AluYb42, AluYb43, AluYb44, AluYb45, AluYb46, AluYb47, AluYb48, AluYb49, AluYb50, AluYb51, AluYb52, AluYb53, AluYb54, AluYb55, AluYb56, AluYb57, AluYb58, AluYb59, AluYb60, AluYb61, AluYb62, AluYb luYbc3a, AluYc1, AluYc2, AluYc5, AluYd2, AluYd3, AluYd3a1, AluYd8, AluYe2, AluYe5, AluYf1, AluYf2, AluYf5, AluYg6, AluYh9, AluYi6, AluYk11, AluYk12 or AluYk13 transposable elements or fragments thereof.

[0122] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) comprises a transposable element or fragment thereof that is derived from or is derived from an AluS series transposable element, e.g., AluSc, AluSc5, AluSc8, AluSg, AluSg1, AluSg4, AluSg7, AluSp, AluSq, AluSq10, AluSq2, AluSq4, AluSx, AluSx3, AluSx4, AluSz, or AluSz6 transposable element.

[0123] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or a fragment thereof that is derived from or from an AluJ series transposable element, e.g., an AluJb, AluJo, AluJr, or AluJr4 transposable element.

[0124] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or fragment thereof that is derived from or is derived from a monomeric Alu-series transposable element, e.g., a FAM, FLAM, FRAM, or PB1D11 transposable element.

[0125] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) comprises a transposable element or fragment thereof derived from or derived from a SINE class transposable element of the SINE2 / tRNA group, e.g., MIR, MIR3, MIRb, MIRc, THER1, THER2, MARE3, UCON3, UCON55, LFSINE_Vert, LmeSINE1b, LmeSINE1c, or MamSINE1 transposable element.

[0126] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or a fragment thereof that is derived from or from an LTR retrotransposon transposable element.

[0127] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) comprises a transposable element or a fragment thereof that is derived from or from an endogenous retroviral (ERV) transposable element.

[0128] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) is selected from the group consisting of ERV1 superfamily transposable elements, e.g., LTR06, LTR9, LTR24C, LTR26, LTR26B, LTR26C, LTR26D, LTR26E, LTR27, LTR27B, LTR27C, LTR29, LTR31, LTR34, LTR36, LTR44, LTR45, LTR45B, LTR45C, LTR48, LTR48B, LTR51, LTR54, LTR54B, LTR56, LTR59, LTR60, LTR60B, LTR64, LTR65, LTR68, LTR70, LTR72, LTR73, LTR74, LTR75, LTR76, LTR77, LTR78, LTR79, LTR80, LTR81, LTR82, LTR83, LTR84, LTR85, LTR86, LTR87, LTR88, LTR89, LTR90, LTR91, LTR92, LTR93, LTR94, LTR95, LTR96, LTR97, LTR98, LTR9 ... and / or MER72B, LTR75_1, LTR78, LTR78B, LTR81A, LTR81AB, LTR81B, LTR81C, LTR2752, MER31A, MER31B, MER34A, MER34A1, MER34C, MER34C2, MER34D, MER39, MER39B, MER49, MER50B, MER50C, MER66A, MER66B, MER66D, MER72B, MER87, MER87B, MER88, MER90, MER90a_LTR (MER90a), MER92A, MER92B, MER92C, MER93, MER95, or MER101B transposable element or a fragment thereof.

[0129] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) comprises a transposable element or a fragment thereof derived from or from an ERV2 superfamily transposable element. In some embodiments, the transposable element is an ERV2 superfamily transposable element of the HML1, HML2, HML3, HML4, HML5, HML6, HML7, HML8, HML9, or HML10 group.

[0130] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or a fragment thereof that is derived from or derived from a MER (Medium reiterated frequency repeat) family member.

[0131] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) is selected from the group consisting of HERV-K14I (HERVK14), LTR14A, LTR14B, HERVK, LTR5, LTR5A, HML3, HERVK9I (HERVK9), MER9a1, MER9a2, MER9a3, HML4, HERVK13I (HERVK13), LTR13, LTR13A, HML5, HERVK22I (HERVK22), LTR22A, LTR22B, LTR22B1, LTR22B2, LTR22C, LTR22C 0, LTR22C2, HML6, HERVK3I, LTR3, LTR3A, LTR3B, HML7, HERVK11DI (HERVK11D), MER11D, HML8, HERVK11I (HERVK11), MER11A, MER11B, MER11C, HML9, HERV-K14CI (HERVK14C), LTR14C, HML10, HERVKC4, LTR14, LTR5B, LTR5_Hs, LTR22, LTR22E, MER9, MER9B, RLTR10B, or RLTR10C transposable elements or fragments thereof.

[0132] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a transposable element or a fragment thereof that is derived from or from an ERV3 superfamily transposable element.

[0133] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) comprises a transposable element or a fragment thereof derived from or derived from an ERV3 superfamily transposable element of the MaLR group, e.g., THE1_I, THE1A, THE1B, THE1C, THE1D, MLT1B, MLT1D, MLT1G, MLT1G1, MLT1G2, MLT1G3, MLT1H1, MLT1H2, MLT1I, MLT1J1, MLT1J2, MLT1K, MLT1L, MLT1M, MLT1N2, or MLT1O transposable element.

[0134] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) is an ERV3 superfamily transposable element of the ERVL series, e.g., HERVL, MLT2A1, MLT2A2, MLT2B3, MLT2C2, MLT2D, MLT2F, ERVL, MLT2B2, ERVL-B4, MLT2B4, ERVL-E, MLT2E, ERV3-16A3_I ERV3-16A3_LTR, including transposable elements or fragments thereof derived from or derived from LTR16A, LTR16A1, LTR16A2, LTR16B, LTR16B1, LTR16B2, LTR16C, LTR16D, LTR16D1, LTR16D2, LTR16E, LTR16E1, LTR16E2, HERV16, LTR16, ERVL47, LTR47B, LTR47B2, LTR47B3, or LTR47B4 transposable elements.

[0135] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an AluSp transposable element or a transposable element that is a derivative thereof.

[0136] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a MER11C transposable element or a transposable element that is a derivative thereof.

[0137] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an AluSx transposable element or a transposable element that is a derivative thereof.

[0138] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an MLT1J transposable element or a transposable element that is a derivative thereof.

[0139] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an AluSg2 transposable element or a transposable element that is a derivative thereof.

[0140] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a THE1A transposable element or a transposable element that is a derivative thereof.

[0141] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an AluJb transposable element or a transposable element that is a derivative thereof.

[0142] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an MTL2B4 or MLT2B4 transposable element or a transposable element that is a derivative thereof.

[0143] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an L2a transposable element or a transposable element that is a derivative thereof.

[0144] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an MLT1J2 transposable element or a transposable element that is a derivative thereof.

[0145] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an AluSq transposable element or a transposable element that is a derivative thereof.

[0146] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an L1MB3 transposable element or a transposable element that is a derivative thereof.

[0147] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (eg, the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises a THE1C transposable element or a transposable element that is a derivative thereof.

[0148] In some embodiments, the synthetic delivery system, polynucleotide, or nucleic acid sequence (e.g., the recognition sequence, the integration sequence, the first nucleic acid sequence, or the second nucleic acid sequence) comprises an AluY transposable element or a transposable element that is a derivative thereof.

[0149] In some embodiments, after contacting, reaching, or internalizing a host cell, the transposable elements disclosed herein utilize the cellular retrotransposition machinery to integrate into the target (e.g., cancer) cell genome. In some embodiments, tissue-specific delivery, uptake, and / or integration can be determined by the sequence of the retrotransposon and host expression of the retrotransposition machinery.

[0150] The synthetic delivery system, polynucleotide, or nucleic acid sequence can include an integration sequence that facilitates integration of the polynucleotide, synthetic delivery system, or cargo into the genome of a target cell. The integration sequence can include, for example, a transposon integration signal. In some embodiments, the synthetic delivery system or polynucleotide lacks an integration sequence.

[0151] In some embodiments, the integration sequence facilitates preferential genomic integration of the ctDNA, synthetic delivery system, polynucleotide, or cargo by target cells compared to control cells. The control cells can be, for example, cells of a different cell type (e.g., a cell type disclosed herein). In some embodiments, the target cells are cancer cells, and the control cells are, for example, non-cancer cells of the same or substantially similar cell type. In some embodiments, the target cells are cancer cells of a first cell type, and the control cells are cancer cells of a second cell type. In some embodiments, the target cells are cancer cells of a first cell type, and the control cells are non-cancer cells of a second cell type.

[0152] The integration sequence may be or include a functional fragment and / or variant of the sequence disclosed herein. For example, the integration sequence may include a subset of nucleotides derived from the nucleotide sequence disclosed herein that facilitates preferential integration into the genome of target cells compared to control cells. In some embodiments, the integration sequence includes a functional fragment and / or variant of the sequence disclosed herein that retains at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the target cell genome integration activity of the nucleotide sequence from which the fragment and / or variant is derived.

[0153] The integration sequence may be a subset of contiguous nucleotides from the sequences disclosed herein, e.g., at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, at least about 200, at least about 210, at least about 220, at least about 230, at least about 240, at least about 250, at least about 260, at least about 270, at least about 280, at least about 290, at least about 300, at least about 310, at least about 320, at least about 330, at least about 340, at least about 350, at least about 360, at least about 370, at least about 380, at least about 390, at least about 400, at least about 410, at least about 420, at least about 430, at least about 440, at least about 450, at least about 460, at least about 470, at least about 480, at least about 490, at least about 500, at least about 510, at least about 520, at least about 530, at least about 540, at least about 550, at least about 560, at least about 570, at least about 580, at least about 590, at least about 600, at least about 610, 0, at least about 200, at most about 10, at most about 20, at most about 30, at most about 40, at most about 50, at most about 60, at most about 70, at most about 80, at most about 90, at most about 100, at most about 110, at most about 120, at most about 130, at most about 140, at most about 150, at most about 160, at most about 170, at most about 180, at most about 190, at most about 200, at most about 250, at most about 300, at most about 350, at most about 400, at most about 450, at most about 500, about 10, about 15, about 20, about 25, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 5 ... 20, about 5-30, about 5-40, about 5-50, about 5-60, about 5-70, about 5-80, about 5-90, about 5-100, about 5-150, about 5-200, about 5-300, about 5-400, about 5-500, about 10-20, about 10-30, about 10-40, about 10-50, about 10-60, about 10 ~70, about 10~80, about 10~90, about 10~100, about 10~150, about 10~200, about 10~300, about 10~400, about 10~500, about 15~20, about 15~30, about 15~40, about 15~50, about 15~60, about 15~70, about 15~80, about 15~90, about 15 ~100, approximately 15~150, approximately 15~200, approximately 15~300, approximately 15~400, approximately 15~500, approximately 20~30, approximately 20~40, approximately 20~50, approximately 20~60, approximately 20~70, approximately 20~80, approximately 20~90, approximately 20~100, approximately 20~150, approximately 20~200, approximately 20~300,Approximately 20-400, approximately 20-500, approximately 30-40, approximately 30-50, approximately 30-60, approximately 30-70, approximately 30-80, approximately 30-90, approximately 30-100, approximately 30-150, approximately 30-200, approximately 30-300, approximately 30-400, approximately 30-500, approximately 40-50, approximately 40-60, approximately 40-70, approximately 40-80, approximately 40-90, approximately 40-100, approximately 40-150, approximately 40-200, approximately 40-300, approximately 40-400, approximately 40-500, approximately 50 It can comprise, consist essentially of, or consist of about 60, about 50 to 70, about 50 to 80, about 50 to 90, about 50 to 100, about 50 to 150, about 50 to 200, about 50 to 300, about 50 to 400, about 50 to 500, about 100 to 150, about 100 to 200, about 100 to 300, about 100 to 400, about 100 to 500, about 200 to 300, about 200 to 400, or about 200 to 500 consecutive nucleotides.

[0154] An integrated sequence can comprise, consist essentially of, or consist of a nucleotide sequence that has at least a minimum percentage sequence identity to a subset of contiguous nucleotides from a sequence disclosed herein.

[0155] In some embodiments, the integration sequence comprises at least 70% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0156] In some embodiments, the integration sequence comprises at least 80% sequence identity for at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0157] In some embodiments, the integration sequence comprises at least 85% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0158] In some embodiments, the integration sequence comprises at least 90% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0159] In some embodiments, the integration sequence comprises at least 95% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0160] In some embodiments, the integration sequence comprises at least 98% sequence identity to at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190 or at least about 200 consecutive nucleotides from any one of SEQ ID NOs: 1-78.

[0161] In some embodiments, the integration sequence is at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, or at least about 200 nucleotides in length.

[0162] In some embodiments, the integration sequence is at most about 10, at most about 20, at most about 30, at most about 40, at most about 50, at most about 60, at most about 70, at most about 80, at most about 90, at most about 100, at most about 110, at most about 120, at most about 130, at most about 140, at most about 150, at most about 160, at most about 170, at most about 180, at most about 190, at most about 200, at most about 250, at most about 300, at most about 350, at most about 400, at most about 450, or at most about 500 nucleotides in length.

[0163] In some embodiments, the integration sequence is about 10, about 15, about 20, about 25, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 250, about 300, about 350, about 400, about 450 or about 500 nucleotides in length.

[0164] In some embodiments, the integration sequence is about 5 to 10, about 5 to 20, about 5 to 30, about 5 to 40, about 5 to 50, about 5 to 60, about 5 to 70, about 5 to 80, about 5 to 90, about 5 to 100, about 5 to 150, about 5 to 200, about 5 to 300, about 5 to 400, about 5 to 500, about 10 to 20, about 10 to 30, about 10 to 40, about 10 to 50, about 10 to 60, about 10 to 70, about 10 to 80, about 10 to 90, about 10 to 100, about 10 to 150, about 10 ~200, about 10~300, about 10~400, about 10~500, about 15~20, about 15~30, about 15~40, about 15~50, about 15~60, about 15~70, about 15~80, about 15~90, about 15~100, about 15~150, about 15~200, about 15~300, about 15~400, about 15~500, about 20~30, about 20~40, about 20~50, about 20~60, about 20~70, about 20~80, about 20~90, about 20~100 , about 20-150, about 20-200, about 20-300, about 20-400, about 20-500, about 30-40, about 30-50, about 30-60, about 30-70, about 30-80, about 30-90, about 30-100, about 30-150, about 30-200, about 30-300, about 30-400, about 30-500, about 40-50, about 40-60, about 40-70, about 40-80, about 40-90, about 40-100, about 40-150, about 40-20 The length is 0, about 40 to 300, about 40 to 400, about 40 to 500, about 50 to 60, about 50 to 70, about 50 to 80, about 50 to 90, about 50 to 100, about 50 to 150, about 50 to 200, about 50 to 300, about 50 to 400, about 50 to 500, about 100 to 150, about 100 to 200, about 100 to 300, about 100 to 400, about 100 to 500, about 200 to 300, about 200 to 400, or about 200 to 500 nucleotides.

[0165] In some embodiments, the integration sequence of the synthetic delivery system or polynucleotide is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, at least 20-fold, at least 21-fold, at least 22-fold, at least 23-fold, at least 24-fold, at least 25-fold, at least 26-fold, at least 27-fold, at least 28-fold, at least 29-fold, at least 30-fold, at least 31-fold, at least 32-fold, at least 33-fold, at least 34-fold, at least 35-fold, at least 36-fold, at least 37-fold, at least 38-fold, at least 39-fold, at least 40-fold, at least 41-fold, at least 42-fold, at least 43-fold, at least 44-fold, at least 45-fold, at least 46-fold, at least 47-fold, at least 48-fold, at least 49-fold, at least 50-fold, at least 51-fold, at least 52-fold, at least 53-fold, at least 54-fold, at least 55-fold, at least 56-fold, at least 57-fold, at 5 fold, at least 20 fold, at least 30 fold, at least 40 fold, at least 50 fold, at least 60 fold, at least 70 fold, at least 80 fold, at least 90 fold, at least 100 fold, at least 150 fold, at least 200 fold, at least 250 fold, at least 300 fold, at least 350 fold, at least 400 fold, at least 500 fold, at least 600 fold, at least 700 fold, at least 800 fold, at least 900 fold, at least 1000 fold, at least 1500 fold, at least 2000 fold or at least 3000 fold greater facilitated genomic integration of the synthetic delivery system, polynucleotide or cargo by the target cell.

[0166] The synthetic delivery system, polynucleotide, or nucleic acid sequence can comprise a recognition sequence and an integration sequence. In some embodiments, the integration sequence and the recognition sequence are coupled, for example, covalently coupled by a phosphodiester bond.

[0167] The synthetic delivery system, polynucleotide, or nucleic acid sequence can include a transposable element (e.g., transposon) sequence or functional fragment thereof. The synthetic delivery system, polynucleotide, or nucleic acid sequence can include two or more transposable element (e.g., transposon) sequences or functional fragments thereof, for example, derived from transposable elements of the same or different origin. The transposable element (e.g., transposon) sequence or functional fragment thereof can be derived from, for example, a class I or class II transposon.

[0168] The polynucleotide can comprise a first nucleic acid sequence identified in, derived from, or from ctDNA, which can originate from a cancer cell of the same, substantially the same, or substantially similar cell type or tissue type as the target cell.

[0169] The first nucleic acid sequence may be present in the 3' region of the polynucleotide. The polynucleotide may further comprise a second nucleic acid sequence, for example, identified in, derived from, or derived from ctDNA. The ctDNA may be derived from or derived from the same, substantially the same, or substantially similar cell type or tissue type as the target cell. The ctDNA may be the same ctDNA from which the first nucleic acid sequence is derived. In some embodiments, the ctDNA is different from the ctDNA from which the first nucleic acid sequence is derived. The second nucleic acid may be present in the 5' region of the polynucleotide. In some embodiments, the first nucleic acid sequence is a recognition sequence disclosed herein, and the second nucleic acid sequence is an integration sequence disclosed herein. In some embodiments, the first nucleic acid sequence is an integration sequence disclosed herein, and the second nucleic acid sequence is a recognition sequence disclosed herein. In some embodiments, the first nucleic acid sequence is a first recognition sequence disclosed herein, and the second nucleic acid sequence is a second recognition sequence disclosed herein.

[0170] The first nucleic acid sequence can be or can comprise a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% sequence identity to any one or more of SEQ ID NOs: 1-78, or a functional fragment thereof as disclosed herein (e.g., having at least a given number of contiguous nucleotides, or having at least a given sequence identity for a given number of contiguous nucleotides, or having retained at least a given percentage of a functional activity as disclosed herein).

[0171] The second nucleic acid sequence can be or can include a nucleic acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% sequence identity to any one or more of SEQ ID NOs: 1-78, or a functional fragment thereof as disclosed herein (e.g., having at least a given number of contiguous nucleotides, or having at least a given sequence identity for a given number of contiguous nucleotides, or having retained at least a given percentage of a functional activity as disclosed herein).

[0172] The first and / or second nucleic acid sequence can comprise a functional fragment (e.g., a recognition sequence or integration sequence) from or derived from any suitable sequence disclosed herein, e.g., any one or more of SEQ ID NOs: 1-78.

[0173] The first nucleic acid sequence may be present in the 5' region of the polynucleotide. The polynucleotide may further comprise a second nucleic acid sequence, for example, identified in, derived from, or derived from ctDNA. The ctDNA may be derived from or derived from the same, substantially the same, or substantially similar cell or tissue type as the target cell. The ctDNA may be the same ctDNA from which the first nucleic acid sequence is derived. In some embodiments, the ctDNA from which the second nucleic acid originates is different from the ctDNA from which the first nucleic acid sequence is derived. The second nucleic acid may be present in the 3' region of the polynucleotide. The first and / or second nucleic acid sequence may comprise a functional fragment (e.g., a recognition sequence or an integration sequence) derived from or derived from any suitable sequence disclosed herein.

[0174] In some embodiments, a portion (e.g., about half) of the nucleic acid sequence may be linked to the 5' end of the nucleic acid cargo, gene, or sequence of interest, and another portion (e.g., about half) of the sequence may be linked to the 3' end of the nucleic acid cargo, gene, or sequence of interest. In some embodiments, the nucleic acid sequence may be linked directly or indirectly to the nucleic acid cargo, gene, or sequence of interest. For example, a first sequence may be linked directly to the 5' end of the nucleic acid sequence containing the nucleic acid cargo, gene, or sequence of interest, and a second sequence may be linked indirectly to the 3' end of the nucleic acid sequence containing the nucleic acid cargo, gene, or sequence of interest. In some embodiments, the compositions of the present disclosure may comprise a guide sequence that is directly linked to the nucleic acid cargo, gene, or sequence of interest at one end (e.g., the 3' end) and directly linked to a recognition sequence or integration sequence at the other end (e.g., the 5' end), thereby indirectly connecting the integration or insertion sequence and the nucleic acid cargo, gene, or sequence of interest.

[0175] In some embodiments, the compositions of the present disclosure comprise a first nucleic acid sequence (e.g., comprising a recognition and / or integration sequence) directly or indirectly linked to one end (e.g., its 3' end) of a nucleic acid cargo, gene, or sequence of interest, and a second nucleic acid sequence (e.g., comprising a recognition and / or integration sequence) directly or indirectly linked to the other end (e.g., its 5' end) of the nucleic acid cargo, gene, or sequence of interest.

[0176] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence (e.g., first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence) comprises a sequence disclosed herein, or a disclosed minimum degree of sequence identity to a sequence disclosed herein, or a fragment thereof. The fragment can be a functional fragment, for example, that acts as a recognition sequence or integration sequence disclosed herein.

[0177] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5%, or about 100% sequence identity to at least 10 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0178] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 15 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0179] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 20 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0180] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5%, or about 100% sequence identity to at least 30 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0181] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 50 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0182] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 75 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0183] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 100 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0184] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 200 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0185] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 300 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0186] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to any one of SEQ ID NOs: 1-78.

[0187] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of, or consists of the nucleotide sequence of any one of SEQ ID NOs: 1-78.

[0188] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:68.

[0189] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:73.

[0190] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:74.

[0191] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:75.

[0192] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:53.

[0193] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:13.

[0194] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:14.

[0195] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of the nucleotide sequence of SEQ ID NO:33.

[0196] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of a nucleic acid sequence having at least about 70%, at least about 80%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to SEQ ID NO:53.

[0197] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:53.

[0198] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 75 contiguous nucleotides of SEQ ID NO:53.

[0199] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises, consists essentially of, or consists of a nucleic acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 95.5%, at least about 96%, at least about 96.5%, at least about 97%, at least about 97.5%, at least about 98%, at least about 98.5%, at least about 99%, at least about 99.5% or about 100% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO:53.

[0200] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises, consists essentially of or consists of a nucleic acid sequence having at most about 70%, at most about 75%, at most about 80%, at most about 85%, at most about 90%, at most about 95%, at most about 97.5%, or at most about 99% sequence identity to any one of SEQ ID NOs: 1-78.

[0201] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises a nucleic acid sequence having one or more nucleotide insertions, deletions, or substitutions compared to any one of SEQ ID NOs: 1-78.

[0202] For example, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence can include at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45 or at least 50 nucleotide insertions compared to any one of SEQ ID NOs: 1-78.

[0203] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises at most 1, at most 2, at most 3, at most 4, at most 5, at most 6, at most 7, at most 8, at most 9, at most 10, at most 11, at most 12, at most 13, at most 14, at most 15, at most 16, at most 17, at most 18, at most 19, at most 20, at most 25, at most 30, at most 35, at most 40, at most 45, or at most 50 nucleotide insertions relative to any one of SEQ ID NOs: 1-78.

[0204] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45 or 50 nucleotide insertions compared to any one of SEQ ID NOs: 1-78.

[0205] The insertion(s) may be at the 5' end, the 3' end, within the sequence, or a combination thereof. The insertion(s) may be adjacent, non-adjacent, or a combination thereof.

[0206] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, at least 400, at least 500, at least 600 or at least 700 nucleotide deletions compared to any one of SEQ ID NOs: 1-78.

[0207] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises at most 1, at most 2, at most 3, at most 4, at most 5, at most 6, at most 7, at most 8, at most 9, at most 10, at most 11, at most 12, at most 13, at most 14, at most 15, at most 16, at most 17, at most 18, at most 19, at most 20, at most 25, at most 30, at most 35, at most 40, at most 45, at most 50, at most 75, at most 100, at most 125, at most 150, at most 175, at most 200, at most 250, at most 300, at most 400, at most 500, at most 600, or at most 700 nucleotide deletions relative to any one of SEQ ID NOs: 1-78.

[0208] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises a 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45 or 50 nucleotide deletion compared to any one of SEQ ID NOs: 1-78.

[0209] The deletion(s) may be at the 5' end, the 3' end, within the sequence, or a combination thereof. The deletion(s) may be contiguous, non-contiguous, or a combination thereof.

[0210] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45 or at least 50 nucleotide substitutions compared to any one of SEQ ID NOs: 1-78.

[0211] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises at most 1, at most 2, at most 3, at most 4, at most 5, at most 6, at most 7, at most 8, at most 9, at most 10, at most 11, at most 12, at most 13, at most 14, at most 15, at most 16, at most 17, at most 18, at most 19, at most 20, at most 25, at most 30, at most 35, at most 40, at most 45, or at most 50 nucleotide substitutions compared to any one of SEQ ID NOs: 1-78.

[0212] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence or integration sequence comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45 or 50 nucleotide substitutions compared to any one of SEQ ID NOs: 1-78.

[0213] The one or more substitutions may be at the 5' end, the 3' end, within the sequence, or a combination thereof. The one or more substitutions may be adjacent, non-adjacent, or a combination thereof.

[0214] The polynucleotides or nucleic acid sequences disclosed herein (e.g., recognition sequences, integration sequences, first nucleic acid sequences, or second nucleic acid sequences) can be from about 10 base pairs (bp) to about 1,000 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be from about 100 base pairs (bp) to about 900 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be from about 200 bp to about 800 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be from about 300 bp to about 700 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be from about 400 bp to about 600 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be at least about 50 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be at least about 100 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be at least about 200 bp in length. The polynucleotides or nucleic acid sequences disclosed herein can be at least about 300 bp in length. The polynucleotide or nucleic acid sequences disclosed herein may be at least about 400 bp in length. The polynucleotide or nucleic acid sequences disclosed herein may be at least about 500 bp in length. The polynucleotide or nucleic acid sequences disclosed herein may be at least about 100 bp in length.

[0215] The polynucleotide or nucleic acid sequences disclosed herein can be at most about 50 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 100 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 200 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 300 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 400 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 500 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 1000 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 5000 bp in length. The polynucleotide or nucleic acid sequences disclosed herein can be at most about 10,000 bp in length.

[0216] In some embodiments, the polynucleotides or nucleic acid sequences disclosed herein (e.g., recognition sequences, integration sequences, first nucleic acid sequences, or second nucleic acid sequences) are at least 10 base pairs in length. In some embodiments, the polynucleotides or nucleic acid sequences disclosed herein are at least 20 base pairs in length. In some embodiments, the polynucleotides or nucleic acid sequences disclosed herein are at least 50 base pairs in length. In some embodiments, the polynucleotides or nucleic acid sequences disclosed herein are at least 100 base pairs in length. In some embodiments, the polynucleotides or nucleic acid sequences disclosed herein are at least 200 base pairs in length. In some cases, the polynucleotides or nucleic acid sequences disclosed herein may be at least 400 base pairs in length.

[0217] In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein (e.g., a recognition sequence, integration sequence, first nucleic acid sequence, or second nucleic acid sequence) is about 10 base pairs to about 100 base pairs in length. In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein can be about 10 base pairs to about 200 base pairs in length. In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein can be about 20 base pairs to about 100 base pairs in length. In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein can be about 100 base pairs to about 200 base pairs in length. In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein can be about 100 base pairs to about 500 base pairs in length. In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein can be about 200 base pairs to about 800 base pairs in length. In some embodiments, a polynucleotide or nucleic acid sequence disclosed herein can be about 400 base pairs to about 2,000 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 400 base pairs to about 5,000 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 1,500 base pairs to about 7,200 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 1,900 base pairs to about 5,800 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 2,000 base pairs to about 10,000 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 5,000 base pairs to about 15,000 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 600 base pairs to about 16,900 base pairs in length. In some embodiments, the polynucleotide or nucleic acid sequence disclosed herein can be from about 8,000 base pairs to about 18,000 base pairs in length.In some embodiments, the polynucleotide or nucleic acid sequences disclosed herein can be from about 10,000 base pairs to about 20,000 base pairs in length, hi some embodiments, the polynucleotide or nucleic acid sequences disclosed herein can be from about 400 base pairs to about 20,000 base pairs in length.

[0218] In some embodiments, the synthetic delivery system, polynucleotide, nucleic acid sequence, first nucleic acid sequence, second nucleic acid sequence, recognition sequence, or integration sequence comprises from about 1 to about 20 insertions, deletions, and / or substitutions compared to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide or nucleic acid sequence comprises from about 20 to about 100 insertions, deletions, and / or substitutions compared to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide or nucleic acid sequence comprises from about 50 to about 250 insertions, deletions, and / or substitutions compared to any one of SEQ ID NOs: 1-78. In some embodiments, the polynucleotide or nucleic acid sequence comprises from about 150 to about 500 insertions, deletions, and / or substitutions compared to any one of SEQ ID NOs: 1-78.

[0219] In some embodiments, the synthetic delivery systems of the present disclosure can be functional, e.g., as "naked" DNA, without requiring a commonly used delivery vector or delivery system, e.g., in some embodiments, the synthetic delivery systems do not require or utilize viral vectors, nanoparticles, lipid nanoparticles, liposomes, exosomes, dendrimers, gene guns, or electroporation.

[0220] In some embodiments, targeted cargo delivery by the synthetic delivery systems disclosed herein can significantly reduce off-target and unwanted adverse effects associated with alternative delivery systems, e.g., non-targeted delivery systems.

[0221] In some embodiments, the synthetic delivery systems of the present disclosure may be combined with or utilize a vector, such as a viral vector, a nanoparticle, a lipid nanoparticle, a liposome, an exosome, or a dendrimer.

[0222] In some embodiments, the candidate polynucleotide for use in the synthetic delivery system comprises a full-length transposable element (e.g., a retrotransposon). In some embodiments, the candidate polynucleotide for use in the synthetic delivery system comprises a high proportion of identified mutations, or two or more identified mutations (e.g., SNVs), for example, favoring a higher number of mutations compared to the reference sequence. Mutations may be shared between ctDNA-target cell co-culture conditions and ctDNA alone, and are absent or substantially absent in the genome of untreated control cells. In some embodiments, the candidate polynucleotide for use in the synthetic delivery system is not located in an intron of a gene, or if present in an intron, is not near an exon (e.g., at least 3 kb, 5 kb, or 10 kb from the exon).

[0223] Table 1 provides illustrative polynucleotide sequences that can be used in the synthetic delivery systems disclosed herein, e.g., for delivery to cancer cells, e.g., hematological cancer cells, e.g., multiple myeloma cells, B cells and / or plasma cells. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7]

[0224] Table 2 provides certain details of illustrative polynucleotides that can be used in the synthetic delivery systems disclosed herein, including, for example, the number of identified genomic insertion sites and / or single nucleotide variants (SNVs or SNPs) identified in the experiments disclosed herein. Certain provided polynucleotides can correspond to the sequences provided in Table 1 and can be used in the synthetic delivery systems disclosed herein, for example, for delivery to cancer cells, e.g., hematological cancer cells, e.g., multiple myeloma cells, B cells, and / or plasma cells. [Table 2-1] [Table 2-2]

[0225] Table 3 provides illustrative polynucleotide sequences that can be used in the synthetic delivery systems disclosed herein, for example, for delivery to cancer cells, eg, pancreatic cancer cells. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11] [Table 3-12] [Table 3-13] [Table 3-14] [Table 3-15] [Table 3-16]

[0226] Table 4 provides certain details of illustrative polynucleotides that may be used in the synthetic delivery systems disclosed herein, including, for example, the number of identified genomic insertion sites and / or single base variants (SNVs or SNPs) identified in the experiments disclosed herein. Certain provided polynucleotides can correspond to the sequences provided in Table 3 and can be used in the synthetic delivery systems disclosed herein, for example, for delivery to cancer cells, e.g., pancreatic cancer cells. [Table 4-1] [Table 4-2] [Table 4-3]

[0227] In some embodiments, the synthetic delivery systems disclosed herein elicit no, substantially no, or a reduced immunogenic response compared to alternative delivery systems, e.g., viral vectors.

[0228] In some embodiments, the synthetic delivery systems disclosed herein comprise a conjugation moiety. The conjugation moiety may be attached to the 5' end, the 3' end, or an internal site along the length of the polynucleotide. The conjugation moiety may be a nucleotide analog (such as bromodeoxyuridine). The conjugation moiety may be a conjugate functional group. The conjugate functional group may be an azide group or an alkyne group. The conjugation moiety may be a modified nucleotide that facilitates conjugation to a chemical entity. The conjugation moiety may be a modified nucleotide that includes a functional group that can be conjugated to a chemical entity. In some embodiments, the synthetic delivery systems disclosed herein comprise multiple conjugation moieties. In some embodiments, the multiple conjugation moieties are the same. In some embodiments, the multiple conjugation moieties are different. In some embodiments, the synthetic delivery system or polynucleotide comprises a first conjugation moiety and a second conjugation moiety.

[0229] The conjugation moiety can be or include a single optimally modified nucleotide (e.g., a modified A, C, G, U, or T containing an azide at the 2' position) that is site-specifically incorporated under optimized conditions (e.g., by solid-phase chemical synthesis). The conjugation moiety can be or include multiple nucleotides containing an azide at the 2' position that are incorporated, for example, by substituting nucleotides during an in vitro transcription reaction (e.g., substituting 5-azido-C3-UTP for UTP). Non-limiting examples of conjugation moieties include modified UTP analogs, 5-azidomethyl-UTP, 5-azido-C3-UTP, 5-azido-PEG4-UTP, 5-ethynyl-UTP, DBCO-PEG4-UTP, vinyl-UTP, 8-azido-ATP, 3'-azido-2',3'-ddATP, 5-azido-PEG4-CTP, 5-DBCO-PEG4-CTP, N6-azidohexyl-3'-dATP, 5-azidopropyl-UTP, and 5-DBCO-PEG4-dCpG.

[0230] The conjugation moiety can be further derivatized by chemical reactions such as click chemistry or any other bioconjugation reaction. The conjugation moiety can be conjugated to a chemical entity by chemical reactions, for example, using click chemistry or the Staudinger reaction. Click chemistry can involve the copper(I)-catalyzed [3+2]-Husgen 1,3-dipolar cycloaddition of an alkyne and an azide to yield a 1,2,3-triazole. Click chemistry can be a copper-free variant of the reaction. Click chemistry can be the inverse electron-demand Diels-Alder reaction between a trans-cyclooctadiene and a tetrazine.

[0231] Polynucleotides can be conjugated to conjugation partners (e.g., cargo) using copper-catalyzed click reactions, such as copper-catalyzed 1,3-dipolar azide-alkyne cycloaddition (CuAAC) of an alkyne-functionalized conjugation partner (e.g., cargo, such as a small molecule) and an azide-functionalized nucleic acid. Linear polynucleotides can be modified at their 3' ends with azide-derivatized nucleotides using poly(A) polymerase. Azides can be conjugated to conjugation partners via copper-catalyzed or strain-promoted azide-alkyne click reactions.

[0232] Polynucleotides can be conjugated to a conjugation partner (e.g., cargo) using the Staudinger reaction. For example, a polynucleotide containing an azide-functionalized nucleotide(s) can be conjugated to an alkyne-functionalized small molecule in the presence of triphenylphosphine-3,3',3"-trisulfonic acid (TPPTS).

[0233] Polynucleotides can be conjugated to conjugation partners (e.g., cargo) using the Suzuki-Miyaura reaction. For example, polynucleotides containing halogenated nucleotide analogs can be subjected to the Suzuki-Miyaura reaction in the presence of a cognate reactive conjugation partner (e.g., cargo). Polynucleotides containing 5-iodouridine triphosphate (IUTP) can be functionalized to iodouridine-labeled polynucleotides in the presence of various suitable boronic acid and ester substrates using, for example, a catalytic system with Pd(OAc)2 and 2-aminopyrimidine-4,6-diol (ADHP) or dimethylamino-substituted ADHP (DMADHP). In another example, polynucleotides containing 8-bromoguanosine can be reacted with arylboronic acids in the presence of a catalytic system made of Pd(OAc)2 and a water-soluble triphenylphosphane-3,3',3"-trisulfonate ligand.

[0234] Polynucleotide conjugates can be produced using Michael addition, for example, by reaction of an electron-rich Michael donor with an α,β-unsaturated compound (Michael acceptor).

[0235] In some embodiments, the conjugation moiety, as used herein, can comprise a hapten group. The hapten group can comprise digoxigenin, 2,4-dinitrophenyl, biotin, avidin, or can be selected from azoles, nitroaryl compounds, benzofurazans, triterpenes, ureas, thioureas, rotenone, oxazoles, thiazoles, coumarins, cyclolignans, heterobiaryl compounds, azoaryl compounds, or benzodiazepines. The hapten group can comprise biotin. The nucleic acid comprising the conjugate moiety can be further linked to, for example, a second nucleic acid, a fluorescent moiety (such as a quantum dot, a dye, or the like), or a conjugate partner, for example, a polymer (such as PEG), a macromolecule (such as a carbohydrate, lipid, polypeptide, or the like).

[0236] In some embodiments, the synthetic delivery systems disclosed herein comprise a polynucleotide conjugated to a cargo, for example, a chemical (e.g., a small molecule), an antibody or fragment thereof, a peptide, a protein, an aptamer, a drug, a second polynucleotide, or a combination thereof.

[0237] In some embodiments, the polynucleotide is conjugated to the conjugation partner (e.g., cargo) using a chelator.

[0238] In some embodiments, the synthetic delivery systems disclosed herein include a first conjugation moiety conjugated to a first conjugation partner (e.g., cargo) and a second conjugation moiety conjugated to a second conjugation partner (e.g., the same or a different conjugation partner and / or cargo). II. Cargo

[0239] In some embodiments, the present disclosure provides compositions and methods for cell-, tissue-, and / or organ-specific binding, targeting, uptake, nuclear localization, and / or genomic integration of cargo. The synthetic delivery systems disclosed herein can be used to deliver cargo to target cells. For example, the recognition sequence of the synthetic delivery system can bind to a component (e.g., a receptor) of the target cell, thereby facilitating binding, uptake, nuclear localization, and / or genomic integration of the cargo of the synthetic delivery system.

[0240] The cargo may be delivered to the membrane or surface of the target cell. The cargo may be delivered to the cytoplasm of the target cell. The cargo may be delivered to the cytosol of the target cell. The cargo may be delivered to the nucleus of the target cell.

[0241] Cargo can be, or comprise, or consist essentially of, or consist of nucleic acid cargo.Cargo can be or comprise DNA, such as dsDNA or ssDNA.Cargo can be, comprise, or encode RNA, such as mRNA, rRNA, tRNA, siRNA, shRNA, or ncRNA.Cargo can be or comprise a transgene.

[0242] The cargo may be or comprise a nucleic acid encoding a protein. The cargo may be or comprise a nucleic acid encoding a therapeutic protein. The cargo may be or comprise a nucleic acid encoding, for example, a cytotoxic polypeptide (e.g., a caspase, e.g., an autoactivating, inducible, or non-inducible caspase, a protein that induces p53-mediated apoptotic cell death, CASP3, CASP8, CASP9, BAX, DNA fragmentation factor (DFF)40, or herpes simplex virus thymidine kinase (HSV-TK)), a tumor suppressor, an antigenic peptide, an antibody or an antigen-binding fragment thereof, an enzyme, a cytokine, a signaling molecule, a microbial (e.g., bacterial or viral) antigen or epitope, a polypeptide that increases an immune response, or a polypeptide that reduces an immune response. The cargo may be or comprise a nucleic acid encoding a diagnostic protein. The cargo may be or comprise a diagnostic protein. The cargo may be, comprise, or encode an immunomodulator. The cargo can be, contain, or encode an immune checkpoint modulator, such as an immune checkpoint inhibitor or activator. The cargo can be, contain, or encode one or more neoantigens or tumor-associated antigens. The cargo can be, contain, or encode a factor that alters the sensitivity of target cells to drugs. The cargo can be a nucleic acid encoding an immune receptor, such as a chimeric antigen receptor (CAR).

[0243] The cargo gene encoding the protein can be operably linked to and / or under the regulatory control of a promoter, such as a promoter that is also part of the cargo. The promoter can be, for example, constitutive, inducible, and / or cell-type specific for expression of RNA and / or protein in target cells. The cargo can be or include a nucleic acid that does not encode a protein.

[0244] In some embodiments, the cargo encodes an endogenous protein or functional RNA molecule that is missing or has impaired function in the subject. In some embodiments, the subject has a mutation in a gene, such as a cancerous mutation, a loss-of-function mutation, or a gain-of-function mutation, that can be treated by providing a therapeutic cargo, for example, a transgene that encodes the missing or impaired protein or RNA, or that inhibits the pathogenic protein or RNA.

[0245] In some embodiments, the cargo is a nucleic acid that inserts into the genome and causes genetic or genomic instability and / or cell death (e.g., of a cancer target cell).

[0246] In some embodiments, the cargo is a nucleic acid comprising natural or unnatural nucleotide analogs or bases.

[0247] The cargo may be or may comprise a polypeptide. The polypeptide may be a therapeutic polypeptide. The polypeptide may be a diagnostic polypeptide. The polypeptide may be an anti-cancer polypeptide. The polypeptide may be a cytotoxic peptide for inducing cell death of a target cell, such as apoptosis, necroptosis, or necrosis. The cargo may be or may comprise, for example, a tumor suppressor, an antigenic peptide, an antibody or antigen-binding fragment thereof, an enzyme, a cytokine, a signaling molecule, a polypeptide that increases an immune response, or a polypeptide that reduces an immune response.

[0248] The cargo may be or include a small molecule or organic molecule (e.g., a small molecule therapeutic or fluorescent dye). The small molecule or organic molecule may be or include a therapeutic agent. The small molecule or organic molecule may be or include a diagnostic agent. The small molecule or organic molecule may be or include a cytotoxic agent. In some embodiments, the small molecule or organic molecule is or includes a radiolabeled or radioactive cytotoxic molecule. The small molecule or organic molecule may be an anti-cancer drug. The small molecule or organic molecule may be a kinase inhibitor. The small molecule may be, for example, a compound (e.g., an organic compound) having a molecular weight of at most 500, at most 600, at most 700, at most 800, at most 900, at most 1000, at most 1100, at most 1200, or at most 1500 daltons.

[0249] The cargo may be or may comprise a therapeutic agent. Non-limiting examples of therapeutic agents that the cargo may be or may comprise include vaccines (e.g., mRNA vaccines), AKT inhibitors, alkylating agents, anti-angiogenic agents, antibiotics, antifolates, antihormonal therapies, anti-inflammatory agents, antimetabolites, anti-VEGF agents, pro-apoptotic agents, aromatase inhibitors, ATM modulators, biologic agents, BRAF inhibitors, BTK inhibitors, chimeric antigen receptors (e.g., nucleotides encoding a CAR), CDK inhibitors, cell growth arrest inducers, chemotherapeutic agents, cytokines, cytotoxic drugs, demethylating agents, differentiation inducers, estrogen receptor antagonists, gene therapy agents, growth factor inhibitors, growth factor receptor inhibitors, HDAC inhibitors, heat shock protein inhibitors, hematopoietic stem cell transplantation (HSCT), hormones, hydrazines, immune checkpoint inhibitors, immunomodulators, immunosuppressants, kinase inhibitors, KRAS inhibitors, matrix metalloproteinase inhibitors, MEK inhibitors, inhibitors, mitotic inhibitors, mTOR inhibitors, multispecific (e.g., bispecific) immune cell engagers, multispecific (e.g., bispecific) killer cell engagers, multispecific (e.g., bispecific) T cell engagers, nitrogen mustards, oncolytic viruses, oxazaphosphorines, p53 reactivators, plant alkaloids, platinum-based drugs, proteasome inhibitors, purine analogs, purine antagonists, pyrimidine antagonists, radiation therapy, ribonucleotide reductase inhibitors, signal transduction inhibitors, RNA silencing (e.g., RNAi) agents, gene editing agents, CRISPR / Cas systems or components thereof, RNA replacement therapy, protein replacement therapy, gene therapy, antibody-drug conjugates, surgery, taxanes, therapeutic antibodies, topoisomerase inhibitors, tyrosine kinase inhibitors, vinca alkaloids, and nucleic acids encoding any of the foregoing applicable agents. The cargo can be, for example, an anti-cancer therapeutic agent.

[0250] The cargo may be or include a drug. The cargo may be or include a non-cancer therapeutic agent, such as a therapeutic agent for a metabolic disease, an autoimmune disease, a neurological disease, or a degenerative disease. The cargo may be or include, for example, a vaccine (e.g., a cancer vaccine), a drug, an immunotherapy, an immune checkpoint inhibitor, a kinase inhibitor, a small molecule, a chemotherapeutic agent, radiation therapy, a biologic, or any combination thereof.

[0251] A cargo that is or comprises a therapeutic agent can modulate (e.g., increase or decrease) the activity of a target gene (e.g., an aberrantly expressed gene) or a product encoded by the target gene, such as a protein or RNA. A cargo that is or comprises a therapeutic agent can modulate (e.g., increase or decrease) the expression of a target gene (e.g., an aberrantly expressed gene). A cargo that is or comprises a therapeutic agent can modulate (e.g., increase or decrease) the activity of a ligand or receptor for a target gene (e.g., an aberrantly expressed gene). In some embodiments, a cargo that is or comprises a therapeutic agent can alter the gene product of an aberrantly expressed gene, for example, by targeting the gene product, the gene's transcript, or an epigenetic factor that affects the gene's characteristics (e.g., expression). Non-limiting examples include targeting the protein encoded by the gene, reducing the expression level of the gene using gene therapy or RNAi, and using RNA vaccines to establish an immune response.

[0252] In some embodiments, the cargo may be or include a dye (eg, a fluorescent dye), a radioisotope, or an imaging agent for use in in vivo diagnostic imaging.

[0253] In some embodiments, the cargo is a detectable label (e.g., a fluorescent label, a quantum dot, a Quasar dye, or a radioactive label).The detectable label can be a radioactive label (e.g., a radioisotope) that facilitates visualization of the distribution of the constructs disclosed herein by positron emission tomography (PET) or single photon emission computed tomography (SPECT).The detectable moiety can comprise a radioisotope of at least one element.

[0254] The cargo may be or may comprise a radionucleotide. The cargo may be or may comprise a polymer. The cargo may be or may comprise a nanoparticle. In some embodiments, the cargo can cause DNA damage from either the outside or the inside of the target cell.

[0255] The cargo may be covalently or non-covalently attached to the polynucleotide or synthetic delivery system.

[0256] In some embodiments, synthetic delivery system does not contain cargo.For example, in some embodiments, polynucleotide disclosed herein can directly exert its effect in target cell, for example, by inducing cell death after polynucleotide binding, uptake, nuclear transport and / or genome integration.In some embodiments, synthetic delivery system acts as an antagonist to reduce the uptake of ctDNA. III. Target cells

[0257] The compositions and methods disclosed herein can be used to deliver cargo to target cells. For example, the uptake of synthetic delivery systems or cargo by target cells can be induced by contacting the target cells with the synthetic delivery systems and / or polynucleotides disclosed herein. The methods disclosed herein can include delivering cargo to target cells.

[0258] The target cell can be a primary cell. The target cell can be a cancer cell, e.g., a hematological tumor cell or a solid tumor cell. The target cell can be a cell line.

[0259] The target cell may be a eukaryotic cell. The target cell may be a mammalian cell. The target cell may be a human cell.

[0260] The target cell can be a white blood cell, such as a lymphoid or myeloid cell. The target cell can be a plasma cell. The target cell can be a multiple myeloma cell. The target cell can be a B cell. The target cell can be a T cell. The target cell can be a lymphocyte. The target cell can be a monocyte or macrophage. The target cell can be a neoplastic myeloid cell. The target cell can be a neoplastic lymphoid cell. The target cell can be a multiple myeloma cell. The target cell can be a leukemia cell. The target cell can be a lymphoma cell.

[0261] The target cell can be a pancreatic cell. The target cell can be an exocrine pancreatic cell. The target cell can be an endocrine pancreatic cell. The target cell can be a pancreatic cancer cell. The target cell can be a bile duct cell. The target cell can be a gallbladder cell.

[0262] The target cell can be a gastrointestinal cell. The target cell can be an intestinal cell, such as a colon cell. The target cell can be a colorectal cancer cell.

[0263] The target cells can be, for example, lymphoma, leukemia, lung cancer, colon cancer, or brain cancer cells.

[0264] In some embodiments, the target cell is a hematological or hematopoietic cell, hi some embodiments, the target cell is a bone marrow-derived cell.

[0265] In some embodiments, the target cell is a solid tumor cell, a liquid tumor cell, a hematological cancer cell, a leukemia cell, a lymphoma cell, a B cell, a bladder cancer cell, a brain cancer (e.g., astrocytoma, glioblastoma, meningioma, or oligodendroglioma) cell, a breast cancer (e.g., ER+, PR+, HER2+, or triple-negative breast cancer) cell, a bone cancer cell, a cervical cancer cell, a colon cancer cell, a colorectal cancer cell, an esophageal cancer cell, a head and neck cancer cell, a kidney cancer cell, a liver cancer cell, a lung cancer cell, a medullary thyroid cancer cell, an oral cancer cell, a nasal cancer cell, an ovarian cancer (e.g., mucinous, endometrioid, clear cell, or undifferentiated) cell, a pancreatic cancer cell, a renal cancer cell, a skin cancer cell, a gastric cancer cell, a pharyngeal cancer cell, a thyroid cancer cell, a uterine cancer cell, a carcinoma cell, a sarcoma cell, an adenoma cell, an adenocarcinoma cell, a cancer cell of unknown tissue of origin, or a neuroendocrine tumor cell.

[0266] In some embodiments, the synthetic delivery systems, polynucleotides, nucleic acid sequences, first nucleic acid sequences, second nucleic acid sequences, recognition sequences or integration sequences, or functional fragments thereof disclosed herein can specifically or preferentially mediate binding, uptake and / or genomic integration of target cells. For example, in some embodiments, the synthetic delivery systems disclosed herein are at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 50-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 0 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, at least 100 times, at least 150 times, at least 200 times, at least 250 times, at least 300 times, at least 350 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1000 times, at least 1500 times, at least 2000 times, or at least 3000 times more capable of binding to, entering, localizing to the nucleus, and / or integrating into the genome of the target cells disclosed herein. The control cells can be, for example, of a different cell or tissue type. The control cells can be, for example, a non-cancer cell type compared to the target cells, which are cancer cells.

[0267] In some embodiments, specificity for target cells can reduce off-target and unwanted side effects, for example, compared to conventional cargo delivery systems (e.g., nanoparticles or viral vectors). IV. Pharmaceutical Compositions

[0268] The compositions disclosed herein can be pharmaceutical compositions (e.g., formulations), and the methods can utilize pharmaceutical compositions. For example, in some embodiments, pharmaceutical compositions are provided that include a compound, such as a synthetic delivery system, a polynucleotide, a cargo, a therapeutic agent, and / or a diagnostic agent, and a pharmaceutically acceptable excipient, carrier, vehicle, or diluent. In some embodiments, the present disclosure provides pharmaceutical compositions that include a synthetic delivery system or a polynucleotide disclosed herein and a pharmaceutically acceptable excipient, carrier, vehicle, or diluent.

[0269] The active agents or compounds described herein (e.g., synthetic delivery systems, polynucleotides, cargos, therapeutic agents, and / or diagnostic agents) can be combined with other chemically and / or pharmaceutically acceptable components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickeners, antioxidants, solubilizers, buffers, osmolytes, salts, surfactants, amino acids, encapsulating agents, bulking agents, cryoprotectants, and / or excipients, to form pharmaceutical compositions. The compositions can facilitate administration of any active agent described herein (e.g., polynucleotides or synthetic delivery systems) to an organism. The compositions can be administered in therapeutically effective amounts as pharmaceutical compositions by a variety of forms and routes, including, for example, intravenous, subcutaneous, intramuscular, rectal, aerosol, parenteral, ocular, pulmonary, transdermal, vaginal, ocular, nasal, oral, sublingual, inhalation, dermal, intrathecal, intranasal, and topical administration (e.g., for use in melanoma). The pharmaceutical compositions can be administered in a local or systemic manner, for example, by injection of the nucleic acid molecules described herein directly into an organ, optionally in a depot.

[0270] Parenteral injections can be formulated for bolus injection or continuous infusion. The pharmaceutical compositions described herein may be in a form suitable for parenteral injection as sterile suspensions, solutions, or emulsions in oily or aqueous vehicles and may contain formulatory agents, such as suspending, stabilizing, and / or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of compounds, such as the synthetic delivery systems, polynucleotides, cargos, therapeutic agents, and / or diagnostic agents described herein, in water-soluble form. Suspensions of the active compound(s) can be prepared as oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. The suspension can also contain suitable stabilizers or agents that increase solubility and / or reduce compound aggregation, allowing for the preparation of highly concentrated solutions. Alternatively or additionally, the compound can be lyophilized or in powder form, for reconstitution with a suitable vehicle, for example, sterile pyrogen-free water, before use.In some cases, compounds such as synthetic delivery systems, polynucleotides, cargos, therapeutic agents and / or diagnostic agents are directly injected into tissue.Compounds such as synthetic delivery systems, polynucleotides, cargos, therapeutic agents and / or diagnostic agents described herein can home to, target, migrate, and penetrate into organs, for example, pancreas, or cells, for example, multiple myeloma cells, when administered to a subject.

[0271] The disclosed compounds, such as synthetic delivery systems, polynucleotides, cargos, therapeutic agents, and / or diagnostic agents, can be applied directly to organs or organ tissues or cells, e.g., the lung or pancreas, during surgical procedures. The disclosed compounds, such as synthetic delivery systems, polynucleotides, cargos, therapeutic agents, and / or diagnostic agents, can be administered locally and formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, and ointments. Such pharmaceutical compositions can contain solubilizers, stabilizers, osmolality enhancers, buffers, and preservatives.

[0272] In practicing the methods of treatment or use provided herein, a therapeutically effective amount of a compound, such as a synthetic delivery system, polynucleotide, cargo, therapeutic agent, and / or diagnostic agent described herein, can be administered in a pharmaceutical composition to a subject having a disease or condition (e.g., cancer) to be treated. In some embodiments, the subject is a mammal, e.g., a human. The therapeutically effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors.

[0273] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers, including excipients and auxiliary agents, that can facilitate the processing of active compounds into preparations that can be used as pharmaceuticals. Formulations can be modified and adapted depending on the chosen route of administration. Pharmaceutical compositions containing the polynucleotides or synthetic delivery systems described herein can be produced, for example, by expressing the polynucleotides in recombinant systems, purifying polynucleotides or complexes containing the polynucleotides, lyophilizing nucleic acid molecules, mixing, dissolving, granulating, dragee-making, mashing, emulsifying, encapsulating, entrapping, or compressing processes. Pharmaceutical compositions can include at least one pharmaceutically acceptable carrier, diluent, or excipient and a compound described herein in free base or pharmaceutically acceptable salt form. Pharmaceutical compositions can contain solubilizers, stabilizers, osmotic enhancers, buffers, and preservatives.

[0274] As described herein, methods for preparing compositions comprising the compounds described herein (e.g., synthetic delivery systems, polynucleotides, nucleic acids, or cargo-attached polynucleotides) include formulating the compounds with one or more inert pharmaceutically acceptable excipients or carriers to form solid, semi-solid, or liquid compositions. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, cachets, and suppositories. These compositions may also contain minor amounts of auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharmaceutically acceptable additives.

[0275] Liquid compositions include, for example, solutions in which the compounds disclosed herein are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds. Semisolid compositions include, for example, gels, suspensions, and creams. The compositions can be in the form of liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquid prior to use, or as emulsions.

[0276] The disclosed compositions can optionally contain a pharmaceutically acceptable preservative.

[0277] Non-limiting examples of pharmaceutically acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), each of which is incorporated herein by reference in its entirety.

[0278] Non-limiting examples of pharmaceutically acceptable carriers include saline, Ringer's solution, and dextrose solution. In some embodiments, the pH of the solution can be about 5 to about 8, about 7 to about 8, or about 7 to about 7.5. Yet another carrier includes sustained-release preparations, such as semipermeable matrices of solid hydrophobic polymers containing the compound. The matrices can be in the form of shaped articles, such as films, liposomes, microparticles, or microcapsules.

[0279] The pharmaceutical compositions described herein can be in unit dosage form suitable for single administration of precise dosage.In unit dosage form, the preparation can be divided into unit doses containing appropriate amounts of one or more synthetic delivery systems, antagonists or therapeutic agents.The unit dosage can be in the form of a package containing different amounts of preparation.Non-limiting examples are packaged injection solutions, vials and ampoules.The aqueous suspension compositions disclosed herein can be packaged in single-dose non-reclosable containers.Multiple-dose reclosable containers can be used, for example, with or without preservatives.The preparations for injection disclosed herein can be in unit dosage form, for example, in ampoules or in multi-dose containers, together with preservatives. V. Method

[0280] In some embodiments, methods for delivering cargo to target cells are disclosed herein. Various assays can be used to demonstrate receptor binding, uptake, nuclear localization, and / or genomic integration of the cargo, synthetic delivery system, or polynucleotide.

[0281] The present disclosure provides methods for the identification, characterization, isolation, synthesis, in vitro testing and in vivo testing of cell- and tissue-type specific synthetic delivery systems.

[0282] The methods disclosed herein can include administering a therapeutic agent to a subject. The subject can be, for example, a subject in need of treatment for a condition to be treated. The therapeutic agent administered to the subject can be or can include, for example, a synthetic delivery system disclosed herein for delivery of a cargo.

[0283] In some embodiments, the methods utilizing the synthetic delivery systems disclosed herein can be used to treat cancer, inflammatory diseases, or autoimmune diseases. In some embodiments, the methods utilizing the synthetic delivery systems to deliver cargo disclosed herein can be used to administer a vaccine, e.g., the cargo can include a vaccine or components thereof, e.g., an antigen and / or an adjuvant or a nucleic acid sequence encoding an antigen and / or an adjuvant.

[0284] The synthetic delivery systems disclosed herein can be used to treat solid tumors. The synthetic delivery systems disclosed herein can be used to treat liquid tumors, such as hematological cancers. The synthetic delivery systems disclosed herein can be used to treat any type of cancer disclosed herein.

[0285] The methods of the present disclosure can be used to treat conditions in a subject in need thereof, such as cancer. The cancer can be a solid tumor or a liquid cancer, such as leukemia or lymphoma. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a hematological cancer, such as a B-cell cancer, such as multiple myeloma. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is colon cancer or colorectal cancer. In some embodiments, the cancer is lung cancer.

[0286] In some embodiments, the cancer comprises bladder cancer, brain cancer (e.g., astrocytoma, glioblastoma, meningioma, or oligodendroglioma), breast cancer (e.g., ER+, PR+, HER2+, or triple-negative breast cancer), bone cancer, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, medullary thyroid cancer, oral cancer, nasal cancer, ovarian cancer (e.g., mucinous, endometrioid, clear cell, or undifferentiated), pancreatic cancer, renal cancer, skin cancer, stomach cancer, pharyngeal cancer, thyroid cancer, or uterine cancer. In some embodiments, the cancer comprises bladder cancer, brain cancer, breast cancer, colon cancer, colorectal cancer, lung cancer, or ovarian cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is brain cancer. In some embodiments, the cancer is breast cancer, e.g., triple-negative breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is bladder cancer.

[0287] In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is a sarcoma. In some embodiments, the cancer is an adenoma. In some embodiments, the cancer is an adenocarcinoma.

[0288] In some embodiments, the cancer is of unknown tissue of origin. In some embodiments, the methods disclosed herein are used to identify the tissue of origin. In some embodiments, the cancer is a neuroendocrine tumor.

[0289] The treatment can include administering a synthetic delivery system to a subject. The treatment can include administering a synthetic delivery system of the present disclosure to a subject intravenously, subcutaneously, intramuscularly, by inhalation, cutaneously, intradermally, topically, orally, sublingually, intrathecally, transdermally, intranasally, intracerebrally, intraspinally, intraarticularly, ocularly, rectally, via a peritoneal route, or directly into the brain, e.g., via an intracerebral ventricular route. The synthetic delivery system can be administered locally. The synthetic delivery system can be administered systemically. The synthetic delivery system can be administered parenterally. The synthetic delivery system or antagonist can be administered by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and / or intestinal mucosa).

[0290] The synthetic delivery system or therapeutic agent described herein can be administered before, during, or after the onset of a disease or condition, and the timing of administering the composition containing the synthetic delivery system or therapeutic agent can vary. For example, the composition can be used as a prophylactic and can be administered continuously to a subject with a condition or a tendency to a disease to reduce the likelihood of the disease or condition occurring. The composition can be administered to a subject already suffering from a disease or condition in an amount sufficient to cure or at least partially halt the symptoms of the disease or condition, or to cure, remedy, improve, or alleviate the condition. The composition can be administered to a subject at the onset of symptoms or as soon as possible thereafter.

[0291] The multiple synthetic delivery systems or therapeutic agents disclosed herein can be administered in any order or simultaneously.When administered simultaneously, the multiple synthetic delivery systems or therapeutic agents can be provided in a single, integrated form or in multiple forms, for example, as multiple separate injections or infusions.The synthetic delivery systems or therapeutic agents can be packaged together or separately in a single package or multiple packages.One or all of the synthetic delivery systems or therapeutic agents can be given in multiple doses.

[0292] In some embodiments, the nucleic acid cargo or component of the polynucleotide is integrated into the target cell genome at a location identified in Table 2, Table 4, Table 7 or Table 8. In some embodiments, the nucleic acid cargo or component of the polynucleotide is integrated into the target cell genome at Chr2:32916224-32916626. In some embodiments, the nucleic acid cargo or component of the polynucleotide is integrated into the target cell genome at Ch16:32628381-32629000.

[0293] The methods and compositions of the present disclosure can be useful for modifying the phenotype of target cell and / or target cell population.In some cases, modifying the phenotype of target cell in a subject can improve the treatment and / or clinical response of the subject to therapeutic intervention.In some embodiments, the methods and compositions of the present disclosure can be used to modify the sensitivity of target cell or target cell population to drugs, for example, to increase the sensitivity of cancer cells to anti-cancer drugs, or to limit or reduce the decrease in the sensitivity of cancer cells to anti-cancer drugs.

[0294] The compositions or methods of the present disclosure can be used, for example, to induce an immune response in a subject after administration of a synthetic delivery system. For example, an immune response can be induced by administering a nucleic acid construct comprising a cargo nucleic acid sequence encoding one or more antigenic or immunogenic peptides or proteins, and such immunogenic peptides or proteins can induce an immune response in a subject after expression. In some embodiments, the synthetic delivery system induces an immune response based on the presence of cytosolic nucleic acid (e.g., DNA). In some embodiments, the synthetic delivery system induces an immune response based on the expression of a transgene from a nucleic acid cargo encoding, for example, a cytokine, antigen, or other immunomodulator.

[0295] In some embodiments, the high specificity and / or efficiency of delivery to target cells (e.g., multiple myeloma cells) by the synthetic delivery systems disclosed herein facilitates their use as therapeutic vectors. For example, in some embodiments, the synthetic delivery systems facilitate preferential delivery to target cells (e.g., hematological cancer cells such as multiple myeloma cells) with less or substantially no delivery to non-target cells (e.g., bone marrow cells that are not hematological cancer cells, multiple myeloma cells, their precursors, cancer stem cells, or combinations thereof).

[0296] In some embodiments, the nucleic acid cargo or component of the polynucleotide is integrated into the genome of the target cell by homologous recombination or homologous recombination repair.In some embodiments, the sequence of the polynucleotide disclosed herein can define the direction of insertion to a substantial extent.In some embodiments, the polynucleotide comprises one or more guide nucleic acid sequences that facilitate insertion at a specific insertion site in the genome of the target cell.

[0297] In some embodiments, the synthetic delivery systems or polynucleotides disclosed herein can be used for diagnostic and monitoring purposes in various chronic, infectious, or inherited (e.g., genetic) diseases, including cancer and certain disorders involving, for example, blood cells (e.g., anemia, thalassemia, hemophilia, or platelet disorders). In some cases, the presence of a cell- and / or tissue-specific recognition sequence disclosed herein can be used as a biomarker for a particular disease or condition and / or to monitor the response to a particular therapeutic intervention (e.g., chemotherapy, targeted therapy, immunotherapy, cell therapy, and / or gene therapy). In some embodiments, the synthetic delivery systems or polynucleotides disclosed herein can be used as diagnostic agents or in diagnostic methods. For example, integration of a polynucleotide, or a portion thereof, into the genome of a cell can be used as a measure or marker to determine the degree of integration of a cargo nucleic acid sequence (e.g., a therapeutic gene sequence) into the genome. In some embodiments, cellular targeting and genomic integration of a polynucleotide, nucleic acid sequence, cargo, or synthetic delivery system described herein, or a component thereof, can be used as a marker for a particular biological effect. In certain instances, the degree of uptake or genomic integration of the cargo, polynucleotide or portion thereof can be a marker or measure of therapeutic efficacy, e.g., cell killing, cargo delivery or expression of the nucleic acid cargo.

[0298] In some embodiments, synthetic delivery systems are used to deliver therapeutic and / or diagnostic cargo to specific cells, cell types, tissues, tissue types or organs of interest.

[0299] In some embodiments, the synthetic delivery systems are used to visualize and / or track a disease or condition (e.g., cancer) in vivo, for example, by delivering a chemical dye (e.g., a fluorescent dye) or a radioisotope to one or more cells associated with the disease or condition. In another example, tumor cells can be visualized and tracked in vivo by delivering, for example, a chemical dye (e.g., a fluorescent dye), a radioisotope, or an imaging agent to a tumor site(s) (e.g., a primary tumor site and / or a metastatic site) using the synthetic delivery systems disclosed herein.

[0300] In some embodiments, the modular nature of the synthetic delivery systems disclosed herein facilitates their use for preventing, treating, and / or diagnosing a variety of diseases and conditions, including chronic, metabolic, and infectious diseases or conditions, e.g., cancer. The methods can include administering an effective amount of the synthetic delivery system, polynucleotide, or pharmaceutical composition to a subject. In some embodiments, an effective amount is an amount sufficient to alleviate (e.g., partially or completely alleviate) one or more signs or symptoms of the disease or condition being treated.

[0301] The synthetic delivery system, polynucleotide or pharmaceutical composition may be administered in combination with one or more other therapeutic agents (e.g., a small molecule drug, an immunotherapeutic agent (e.g., an immune checkpoint blocker), a therapeutic antibody, or any other therapeutic agent disclosed herein).

[0302] Non-limiting examples of therapeutic agents that can be administered to a subject in combination with the synthetic delivery systems or polynucleotides disclosed herein include anti-cancer therapeutic agents, vaccines (e.g., mRNA vaccines), cancer vaccines, AKT inhibitors, alkylating agents, anti-angiogenic agents, antibiotics, antifolates, antihormonal therapies, anti-inflammatory agents, antimetabolites, anti-VEGF agents, pro-apoptotic agents, aromatase inhibitors, ATM modulators, biologic agents, BRAF inhibitors, BTK inhibitors, CAR-T cells, CAR-NK cells, CDK inhibitors, cell growth arrest inducers, cell therapies, chemotherapy, cytokine therapy, cytotoxic drugs, demethylating agents, differentiation inducers, estrogen receptor antagonists, gene therapy agents, growth factor inhibitors, growth factor receptor inhibitors, HDAC inhibitors, heat shock protein inhibitors, hematopoietic stem cell transplantation (HSCT), hormones, hydrazines, immune checkpoint inhibitors (e.g., targeting the PD-1 / PD-L1 axis, or other immune checkpoints, e.g., cytotoxic T-lymphocyte-associated molecule 4 (CTLA-4) or LAG-3, TIM-3, TIGIT, VISTA, or B7 / H3) immune checkpoint inhibitors), immunomodulators, kinase inhibitors, KRAS inhibitors, matrix metalloproteinase inhibitors, MEK inhibitors, mitotic inhibitors, mTOR inhibitors, multispecific (e.g., bispecific) immune cell-engaging factors, multispecific (e.g., bispecific) killer cell-engaging factors, multispecific (e.g., bispecific) T cell-engaging factors, nitrogen mustard, oncolytic viruses, oxazaphosphorines, p53 reactivators, plant alkaloids, platinum-based agents, proteasome inhibitors, purine analogs, purine antagonists The present invention also includes, but is not limited to, therapeutic agents such as agonists, pyrimidine antagonists, radiation therapy, ribonucleotide reductase inhibitors, signal transduction inhibitors, RNA silencing (e.g., RNAi) agents, gene editing agents, CRISPR / Cas systems or components thereof, RNA replacement therapy, protein replacement therapy, gene therapy, antibody drug conjugates, surgery, taxanes, therapeutic antibodies, topoisomerase inhibitors, transgenic T cells, tyrosine kinase inhibitors, vinca alkaloids, and nucleic acids encoding any of the preceding agents, as applicable.

[0303] The methods and compositions of the present disclosure can be used to prevent, treat, arrest, reverse, or alleviate symptoms of a condition. Treatment can include treating a subject (e.g., an individual suffering from a disease or condition, a domestic animal, a wild animal, or an experimental animal) with a synthetic delivery system, polynucleotide, or pharmaceutical composition disclosed herein. The disease can be or include cancer or a tumor. In treating a condition, the delivery system, polynucleotide, or pharmaceutical composition of the present disclosure can be contacted with the tumor or cancerous cells. The subject can be a mammal. The subject can be a human. The subject can be, for example, a human; a non-human primate, such as chimpanzees and other ape and monkey species; livestock, cows, horses, sheep, goats, and pigs; or domestic animals, such as rabbits, dogs, and cats; or experimental animals, including rodents, such as rats, mice, and guinea pigs. In some embodiments, the subject is a non-rodent subject. The subject can be a female subject. The subject can be a male subject. Treatment can be provided to the subject before, at, or after the clinical onset of the disease.

[0304] In some embodiments, disclosed herein are methods for delivering cargo to a target cell. The method can include contacting the target cell or a receptor on the surface of the target cell with a synthetic delivery system disclosed herein that includes a polynucleotide.

[0305] The compositions and methods disclosed herein can utilize receptors. The disclosed methods can include, for example, delivering cargo to a target cell by contacting the receptor with a synthetic delivery system or polynucleotide that binds to the receptor, and / or administering an agent that binds to the receptor and / or induces receptor-mediated uptake. The receptor can be expressed by a target cell as disclosed herein. In some embodiments, receptor expression is specific to the target cell, target cell type, or target cell population, thereby facilitating specific delivery of cargo to the target cell, target cell type, or target cell population. In some embodiments, receptor expression is relatively higher in the target cell, target cell type, or target cell population than in a control cell, control cell type, or control cell population, thereby facilitating delivery of cargo to the target cell, target cell type, or target cell population. In some embodiments, receptor expression is not specific to the target cell, target cell type, or target cell population. Binding of a synthetic delivery system or polynucleotide to a receptor disclosed herein can induce uptake of the synthetic delivery system, polynucleotide or cargo associated therewith by, for example, endocytosis, receptor-mediated endocytosis, clathrin-dependent endocytosis, caveolae-dependent endocytosis, CLIC / GEEC pathway uptake, lipid raft-associated endocytosis, phagocytosis, macropinocytosis, micropinocytosis, pinocytosis or endosomal recycling.

[0306] The receptor may be or comprise an MHC protein or a component thereof. The receptor may be or comprise a human leukocyte antigen or a component thereof. The receptor may be or comprise a class I MHC protein or a component thereof. For example, the receptor may be or comprise an HLA-A or a component thereof. The receptor may be or comprise a protein that interacts with an MHC class I molecule, such as APLP2.

[0307] The receptor may be or include a G protein-coupled receptor (GPCR) or its component. The GPCR may be a class A (rhodopsin-like), class B (e.g., secretin and adhesion subfamily), class C (glutamate), or class F (Frizzled) family or subfamily GPCR. The GPCR may be a GPCR that induces a cyclic adenosine 3,5-monophosphate (cAMP) response, calcium mobilization, or phosphorylation of extracellular-regulated protein kinase 1 / 2 (pERK1 / 2). For example, the receptor may be or include CD97 or a component thereof.

[0308] The receptor may be or may include an adhesion protein or a component thereof. For example, the receptor may be or may include CD97 or a component thereof, PTK7 or a component thereof, ITGB5 or a component thereof, ITGA2B or a component thereof, or αVβ5 integrin or a component thereof.

[0309] The receptor may be or may include an integrin or a component thereof, for example, ITGB5 or a component thereof, ITGA2B or a component thereof, or αVβ5 integrin or a component thereof.

[0310] The receptor may be or comprise a tyrosine phosphatase or a component thereof. The receptor may be or comprise a protein tyrosine phosphatase or a component thereof. The receptor may be or comprise a protein tyrosine phosphatase receptor or a component thereof. The receptor may be or comprise, for example, a PTPRF or a component thereof.

[0311] The receptor may be or include an ion channel or a component thereof. The receptor may include a sensory neuron ion channel, such as a nociceptor ion channel. The receptor may include a function in force sensation. The receptor may be or include, for example, TMEM120A or a component thereof.

[0312] The receptor may be or include a mineral transporter or a component thereof. The receptor may be or include ferroxidase or a component thereof. The receptor may be or include, for example, ceruloplasmin or a component thereof.

[0313] The receptor may be or may include a metalloprotein or a component thereof. The receptor may be or may include, for example, ceruloplasmin or a component thereof.

[0314] The receptor may be or include a nutrient transporter or a component thereof. The receptor may be or include a monocarboxylic acid transporter or a component thereof. The receptor may be or include, for example, SLC16A1 or a component thereof. The receptor may be or include, for example, an amino acid transporter or a component thereof. The receptor may be or include, for example, SLC7A5 or a component thereof.

[0315] The receptor may be or include a receptor for fibronectin, fibrinogen, plasminogen, prothrombin, thrombospondin, vitronectin, or a combination thereof. For example, the receptor may be or include ITGA2B or a component thereof.

[0316] The receptor may be or comprise an ATPase or a component thereof. The receptor may be or comprise a sodium / potassium transporting ATPase or a component thereof. The receptor may be or comprise, for example, ATP1B1 or a component thereof. The receptor may be or comprise a calcium transporter or a component thereof. The receptor may be or comprise an ion transporting ATPase or a component thereof. The receptor may be, for example, ATP2B4 or a component thereof.

[0317] The receptor may be or include, for example, an Armadillo-like protein, including one or more armadillo repeats. The receptor may be or include, for example, a member of the p120(ctn) / plakophilin subfamily of Armadillo-like proteins, including CTNND1, CTNND2, PKP1, PKP2, PKP4, or ARVCF. In some embodiments, the receptor is PKP4.

[0318] The receptor may be or may include an APP (amyloid precursor protein) family member, such as APP, APLP1, or APLP2. In some embodiments, the receptor is APLP2.

[0319] The receptor may be or include a kinase. The receptor may be or include a lipid kinase. The receptor may be or include a phosphatidylinositol kinase. The receptor may be or include a phosphatidylinositol-5-phosphate 4-kinase family member. The receptor may be capable of catalyzing the phosphorylation of phosphatidylinositol-5-phosphate at the fourth hydroxyl of the myo-inositol ring to form phosphatidylinositol-5,4-bisphosphate. The receptor may be or include PIP4K2A.

[0320] The receptor may be or may include P142A or a component thereof.

[0321] In some embodiments, a particular cargo or type of cargo disclosed herein may be delivered by targeting a synthetic delivery system to a receptor or type of receptor disclosed herein, for example, by targeting via a polynucleotide and / or recognition sequence.

[0322] In some embodiments, targeting of a synthetic delivery system to a receptor or type of receptor disclosed herein can increase specific binding, uptake, nuclear localization and / or genomic integration of the synthetic delivery system, polynucleotide and / or cargo by the target cell compared to a control cell.

[0323] In some embodiments, the receptor comprises, consists essentially of, or consists of a single protein, receptor, or subunit disclosed herein. In some embodiments, the receptor comprises, consists essentially of, or consists of two proteins or subunits disclosed herein, or two receptors, proteins, or subunits contribute to uptake. In some embodiments, the receptor comprises, consists essentially of, or consists of three proteins or subunits disclosed herein, or three receptors, proteins, or subunits contribute to uptake.

[0324] The methods disclosed herein can include measuring the expression level of a receptor associated with the uptake of ctDNA, polynucleotides, transposable elements, or synthetic delivery systems. For example, a biological sample can be assayed to determine whether the receptor is expressed in the biological sample at a level associated with the uptake of ctDNA, polynucleotides, transposable elements, or synthetic delivery systems. The assay can measure receptor expression at the protein level and / or mRNA level, e.g., quantify or qualify the expression level of the receptor protein or mRNA encoding the receptor. Any suitable assay can be used to determine the expression level of the receptor. Non-limiting examples of assays that can be used include immunohistochemistry, ELISA, multiplex immunoassay, mass spectrometry (e.g., targeted or non-targeted), fluorescence microscopy, flow cytometry, quantitative PCR (e.g., quantitative reverse transcriptase real-time PCR), digital PCR, and RNA sequencing. In some embodiments, the assay is quantitative. In some embodiments, the assay is semi-quantitative, e.g., providing expression levels compared to a control. The biological sample can include or be a liquid. The biological sample can be a liquid biopsy. A biological sample can be or include, for example, blood (e.g., whole blood). A biological sample can be or include a solid. A biological sample can be or include a solid tissue sample from any organ or tissue. A biological sample can be or include a biopsy containing or suspected of containing tumor tissue. A biological sample can include, for example, tumor tissue of any cancer or tumor type disclosed herein. A biological sample can include, for example, cancer cells of any cancer or tumor type disclosed herein. A biological sample can primarily include cells from a specific organ or from tissue within a specific organ.

[0325] The degree of sequence identity between two sequences can be determined by comparing two sequences using a computer program commonly used for this purpose, such as a global or local alignment algorithm.Non-limiting examples include BLASTp, BLASTn, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, GAP, BESTFIT or other suitable method or algorithm.Needleman and Wunsch global alignment algorithm can be used to align two sequences over their entire length, maximizing the number of matches and minimizing the number of gaps.Default settings can be used. VI. Embodiment

[0326] Embodiment 1. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide, and the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 10 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0327] Embodiment 2. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 15 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0328] Embodiment 3. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 20 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0329] Embodiment 4. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 30 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0330] Embodiment 5. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 50 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0331] Embodiment 6. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 75 contiguous nucleotides of any one of SEQ ID NOs: 1-78.

[0332] Embodiment 7. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to at least 100 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0333] Embodiment 8. The pharmaceutical composition of any one of embodiments 1 to 7, wherein the polynucleotide comprises a nucleotide sequence having at least 80% sequence identity to any one of SEQ ID NOs: 1 to 78.

[0334] Embodiment 9. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0335] Embodiment 10. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 15 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0336] Embodiment 11. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 20 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0337] Embodiment 12. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 30 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0338] Embodiment 13. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 50 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0339] Embodiment 14. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 75 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0340] Embodiment 15. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of any one of SEQ ID NOs: 1-78.

[0341] Embodiment 16. The pharmaceutical composition of any one of the preceding embodiments, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to any one of SEQ ID NOs: 1-78.

[0342] Embodiment 17. The pharmaceutical composition of any one of the preceding embodiments, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to any one of SEQ ID NOs: 1-78.

[0343] Embodiment 18. The pharmaceutical composition of any one of the preceding embodiments, wherein the polynucleotide comprises a nucleotide sequence having at least 97% sequence identity to any one of SEQ ID NOs: 1-78.

[0344] Embodiment 19. The pharmaceutical composition of embodiment 1, wherein the polynucleotide comprises any one of the nucleotide sequences of SEQ ID NOs: 1 to 78.

[0345] Embodiment 20. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide, and the polynucleotide is selected from the group consisting of AluSp, MER11C, AluY, 2L2a, ALluY, ALR / alpha, ALU, AluJb, AluS1, AluSc8, AluSg, ALUSg2, AluSq, AluSq2, ALUSx, AluSx, AluSx1, AluSz, AluSz6, AluYc3, ASLUSq2, ERVK / LTR, ERVL, ERVL-MaLR, FLAM_C, and AluY, HERV17-int, HERV9N-int, L1M1, L1MB3, L1ME4b, L1MEg, L1P1, L1P3, L1PA10, L1PA15, L1PA7, L1PB4, L2a, L2a / LTR40b / MLT1J2, LINE / L1, L INE / L2, LTR, LTR / ERV1, LTR / ERVL, LTR / ERVL-MaLR, LTR / Gypsy, LTR41C, LTR81B, Mam_R4, Mamr4, Many, MER11B, MER41E, MIR, MIRB, MIRc, MIRc- A pharmaceutical composition comprising a transposable element that is part L2, MLT1J2, MLT2B1, MLT1J2, MLT2B4, parAluSp-FULLMTL1J2, REP522, satellite / center, SINE / Alu, SINE / MIR, THE1A, THE1B, THE1C, or Tigger3a transposable element.

[0346] Embodiment 21. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, the recognition sequence facilitating binding or uptake of the synthetic delivery system by a target cell, the recognition sequence comprising a nucleic acid sequence identified in circulating tumor DNA (ctDNA) from a cell type substantially similar to the target cell, and the polynucleotide comprises a short interspersed nucleotide sequence (SINE), a long interspersed nucleotide sequence (LINE), an ERVL or an ERVK transposable element.

[0347] Embodiment 22. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an AluSp transposable element.

[0348] Embodiment 23. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises a MER11C transposable element.

[0349] Embodiment 24. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an AluSx transposable element.

[0350] Embodiment 25. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an MLT1J transposable element.

[0351] Embodiment 26. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an AluSg2 transposable element.

[0352] Embodiment 27. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises a THE1A transposable element.

[0353] Embodiment 28. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an AluJb transposable element.

[0354] Embodiment 29. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an MLT2B4 transposable element.

[0355] Embodiment 30. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an L2a transposable element.

[0356] Embodiment 31. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an MLT1J2 transposable element.

[0357] Embodiment 32. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an AluSq transposable element.

[0358] Embodiment 33. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an L1MB3 transposable element.

[0359] Embodiment 34. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises a THE1C transposable element.

[0360] Embodiment 35. The pharmaceutical composition of any one of embodiments 1 to 21, wherein the polynucleotide comprises an AluY transposable element.

[0361] Embodiment 36 The pharmaceutical composition of any one of the preceding embodiments, wherein the polynucleotide comprises double-stranded DNA.

[0362] Embodiment 37. The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system further comprises a cargo.

[0363] Embodiment 38. The pharmaceutical composition of embodiment 37, wherein the cargo comprises a nucleic acid cargo.

[0364] Embodiment 39. The pharmaceutical composition of embodiment 38, wherein the nucleic acid cargo is attached to the 3' end of the polynucleotide.

[0365] Embodiment 40. The pharmaceutical composition of embodiment 38, wherein the nucleic acid cargo is attached to the 5' end of the polynucleotide.

[0366] Embodiment 41 The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system comprises a promoter.

[0367] Embodiment 42. A pharmaceutical composition according to any one of embodiments 38 to 41, wherein the nucleic acid cargo encodes a tumor suppressor protein.

[0368] Embodiment 43. A pharmaceutical composition according to any one of embodiments 37 to 42, wherein the cargo comprises a cytotoxic cargo.

[0369] Embodiment 44. A pharmaceutical composition according to any one of embodiments 37 to 43, wherein the cargo comprises a therapeutic cargo.

[0370] Embodiment 45. The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system does not utilize a viral vector, a nanoparticle, a lipid nanoparticle, a liposome, an exosome, a dendrimer, a gene gun, or electroporation.

[0371] Embodiment 46. A method of delivering a cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition or synthetic delivery system according to any one of the preceding embodiments.

[0372] Embodiment 47 The method of embodiment 46, wherein the cargo is delivered to the nucleus of the target cell.

[0373] Embodiment 48 The method of embodiment 46 or embodiment 47, wherein the synthetic delivery system further comprises an integration sequence that facilitates integration of the polynucleotide, synthetic delivery system, or cargo into the genome of the target cell.

[0374] Embodiment 49. The method of any one of embodiments 46 to 48, wherein the cargo comprises a nucleic acid cargo.

[0375] Embodiment 50. The method of embodiment 49, wherein the nucleic acid cargo is integrated into the genome of the target cell.

[0376] Embodiment 51 The method of embodiment 49, wherein the nucleic acid cargo is integrated into the genome of the target cell at a location identified in Table 2, Table 4, Table 7, or Table 8.

[0377] Embodiment 52 The method of embodiment 49, wherein the nucleic acid cargo is integrated into the target cell genome at Chr2:32916224-32916626.

[0378] Embodiment 53. The method of embodiment 49, wherein the nucleic acid cargo is integrated into the target cell genome at Ch16:32628381-32629000.

[0379] Embodiment 54. The method of any one of embodiments 46 to 53, wherein the target cells are leukocytes.

[0380] Embodiment 55. The method of any one of embodiments 46 to 53, wherein the target cells are plasma cells.

[0381] Embodiment 56. The method of any one of embodiments 46 to 53, wherein the target cells are cancer cells.

[0382] Embodiment 57. The method of any one of embodiments 46 to 53, wherein the target cells are multiple myeloma cells.

[0383] Embodiment 58. The method of any one of embodiments 46 to 53, wherein the target cells are pancreatic cells.

[0384] Embodiment 59. The method of any one of embodiments 46 to 53, wherein the target cells are pancreatic cancer cells.

[0385] Embodiment 60. The method of any one of embodiments 46 to 53, wherein the target cells are gastrointestinal cells.

[0386] Embodiment 61. The method of any one of embodiments 46 to 53, wherein the target cells are colorectal cancer cells.

[0387] Embodiment 62. A method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described in any one of embodiments 1 to 45.

[0388] Embodiment 63. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell, and wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13 or 14.

[0389] Embodiment 64. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13.

[0390] Embodiment 65. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 13.

[0391] Embodiment 66. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 13.

[0392] Embodiment 67. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 13.

[0393] Embodiment 68. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 13.

[0394] Embodiment 69. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13.

[0395] Embodiment 70. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 13.

[0396] Embodiment 71. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 13.

[0397] Embodiment 72. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 13.

[0398] Embodiment 73. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 13.

[0399] Embodiment 74. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 13.

[0400] Embodiment 75. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 13.

[0401] Embodiment 76. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 13.

[0402] Embodiment 77. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 14.

[0403] Embodiment 78. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 14.

[0404] Embodiment 79. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 14.

[0405] Embodiment 80. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO: 14.

[0406] Embodiment 81. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 14.

[0407] Embodiment 82. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 14.

[0408] Embodiment 83. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 14.

[0409] Embodiment 84. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 14.

[0410] Embodiment 85. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 14.

[0411] Embodiment 86. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 14.

[0412] Embodiment v87. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:14.

[0413] Embodiment 88. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 14.

[0414] Embodiment 89. The pharmaceutical composition of embodiment 63, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 14.

[0415] Embodiment 90. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell, and wherein the recognition sequence comprises (a) a nucleotide sequence having at least 90% sequence identity to at least 60 contiguous nucleotides of SEQ ID NO:33, or (b) a nucleotide sequence having at least 91% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:33.

[0416] Embodiment 91. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 60 consecutive nucleotides of SEQ ID NO: 33.

[0417] Embodiment 92. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 91% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 33.

[0418] Embodiment 93. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 75 consecutive nucleotides of SEQ ID NO: 33.

[0419] Embodiment 94. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 33.

[0420] Embodiment 95. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 33.

[0421] Embodiment 96. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 33.

[0422] Embodiment 97. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 33.

[0423] Embodiment 98. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 33.

[0424] Embodiment 99. The pharmaceutical composition of embodiment 90, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 33.

[0425] Embodiment 100. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, and the recognition sequence comprises an AluSp, AluSx, AluSg2, ERV2 superfamily, or ERV3 superfamily transposable element, or a functional fragment thereof, that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell.

[0426] Embodiment 101. The pharmaceutical composition of embodiment 100, wherein the recognition sequence comprises an AluSp transposable element or a functional fragment thereof.

[0427] Embodiment 102. The pharmaceutical composition of embodiment 100, wherein the recognition sequence comprises an AluSx transposable element or a functional fragment thereof.

[0428] Embodiment 103. The pharmaceutical composition of embodiment 100, wherein the recognition sequence comprises an AluSg2 transposable element or a functional fragment thereof.

[0429] Embodiment 104. The pharmaceutical composition of embodiment 100, wherein the recognition sequence comprises an ERV2 superfamily transposable element or a functional fragment thereof.

[0430] Embodiment 105. A pharmaceutical composition described in embodiment 104, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises an HML8 group transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0431] Embodiment 106. A pharmaceutical composition according to embodiment 104 or 105, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises a MER family transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell.

[0432] Embodiment 107. A pharmaceutical composition described in any one of embodiments 104 to 106, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises a MER11C transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0433] Embodiment 108. The pharmaceutical composition of embodiment 100, wherein the recognition sequence comprises an ERV3 superfamily transposable element or a functional fragment thereof.

[0434] Embodiment 109. The pharmaceutical composition of embodiment 108, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises a MaLR group transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0435] Embodiment 110. A pharmaceutical composition described in embodiment 108 or 109, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises a THE1A transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0436] Embodiment 111. A pharmaceutical composition described in embodiment 108 or 109, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises an MLT1J2 transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell.

[0437] Embodiment 112. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell, and wherein the polynucleotide comprises a nucleotide sequence having at least 86% sequence identity to any one of SEQ ID NOs: 7, 16, 25, 29 and 31.

[0438] Embodiment 113. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:7.

[0439] Embodiment 114. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 16.

[0440] Embodiment 115. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 25.

[0441] Embodiment 116. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 29.

[0442] Embodiment 117. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 31.

[0443] Embodiment 118. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:7.

[0444] Embodiment 119. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 16.

[0445] Embodiment 120. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 25.

[0446] Embodiment 121. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 29.

[0447] Embodiment 122. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 31.

[0448] Embodiment 123. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:7.

[0449] Embodiment 124. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 16.

[0450] Embodiment 125. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 25.

[0451] Embodiment 126. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 29.

[0452] Embodiment 127. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 31.

[0453] Embodiment 128. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 7.

[0454] Embodiment 129. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 16.

[0455] Embodiment 130. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 25.

[0456] Embodiment 131. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 29.

[0457] Embodiment 132. The pharmaceutical composition of embodiment 112, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 31.

[0458] Embodiment 133. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates binding of the synthetic delivery system to a target cell.

[0459] Embodiment 134. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates binding of the synthetic delivery system to target cells that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

[0460] Embodiment 135. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates binding of the synthetic delivery system to target cells that is at least 50% higher compared to control cells of a different cell type.

[0461] Embodiment 136. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates uptake of the synthetic delivery system or a component thereof by a target cell.

[0462] Embodiment 137. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates uptake of the synthetic delivery system by target cells that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

[0463] Embodiment 138. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates uptake of the synthetic delivery system by target cells that is at least 50% higher compared to control cells of a different cell type.

[0464] Embodiment 139. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates nuclear localization of the synthetic delivery system within the target cell that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

[0465] Embodiment 140. The pharmaceutical composition of any one of the preceding embodiments, wherein the recognition sequence facilitates nuclear localization of the synthetic delivery system in target cells that is at least 50% higher compared to control cells of a different cell type.

[0466] Embodiment 141. A pharmaceutical composition according to any one of the preceding embodiments, wherein the recognition sequence facilitates binding of the synthetic delivery system to and uptake of the synthetic delivery system or a component thereof by a target cell.

[0467] Embodiment 142. The pharmaceutical composition of any one of the preceding embodiments, wherein the recognition sequence comprises a nucleic acid sequence identified in circulating tumor DNA (ctDNA) from a cell type substantially similar to the target cell.

[0468] Embodiment 143. The pharmaceutical composition of any one of the preceding embodiments, wherein the polynucleotide comprises double-stranded DNA.

[0469] Embodiment 144. The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system further comprises a cargo.

[0470] Embodiment 145. The pharmaceutical composition of embodiment 144, wherein the cargo comprises an anti-cancer therapeutic agent.

[0471] Embodiment 146. A pharmaceutical composition according to embodiment 144 or 145, wherein the cargo comprises a nucleic acid cargo.

[0472] Embodiment 147. The pharmaceutical composition of embodiment 147, wherein the nucleic acid cargo encodes a tumor suppressor protein.

[0473] Embodiment 148. A pharmaceutical composition according to any one of embodiments 144 to 147, wherein the cargo is attached to the 3' end of the polynucleotide.

[0474] Embodiment 149. A pharmaceutical composition according to any one of embodiments 144 to 148, wherein the cargo is attached to the 5' end of the polynucleotide.

[0475] Embodiment 150. A pharmaceutical composition according to any one of embodiments 144 to 149, wherein the cargo comprises a cytotoxic cargo.

[0476] Embodiment 151. A pharmaceutical composition according to any one of embodiments 144 to 150, wherein the cargo comprises a therapeutic cargo.

[0477] Embodiment 152. A pharmaceutical composition according to any one of embodiments 144 to 151, wherein the cargo comprises a polypeptide.

[0478] Embodiment 153. A pharmaceutical composition according to any one of embodiments 144 to 152, wherein the cargo comprises a small molecule.

[0479] Embodiment 154. The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system further comprises a promoter.

[0480] Embodiment 155. The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system further comprises an integration sequence that facilitates integration of the polynucleotide, synthetic delivery system, cargo or a component thereof into the genome of the target cell.

[0481] Embodiment 156. The pharmaceutical composition of embodiment 155, wherein the integration sequence facilitates genomic integration of the synthetic delivery system, polynucleotide, cargo or a component thereof in the target cell that is at least 50% higher compared to a control synthetic delivery system lacking the integration sequence.

[0482] Embodiment 157. A pharmaceutical composition according to any one of embodiments 155 to 156, which facilitates genomic integration of the synthetic delivery system, polynucleotide, cargo or a component thereof by the target cell, wherein the integration sequence is at least 50% higher compared to control cells of a different cell type.

[0483] Embodiment 158. The pharmaceutical composition of any one of the preceding embodiments, wherein the synthetic delivery system does not utilize a viral vector, a nanoparticle, a lipid nanoparticle, a liposome, an exosome, a dendrimer, a gene gun, or electroporation for delivery to the target cell.

[0484] Embodiment 159. A pharmaceutical composition according to any one of embodiments 62 to 158, wherein the target cells are leukocytes.

[0485] Embodiment 160. A pharmaceutical composition according to any one of embodiments 62 to 159, wherein the target cells are plasma cells.

[0486] Embodiment 161. A pharmaceutical composition according to any one of embodiments 62 to 160, wherein the target cells are cancer cells.

[0487] Embodiment 162. A pharmaceutical composition according to any one of embodiments 62 to 161, wherein the target cells are multiple myeloma cells.

[0488] Embodiment 163. A pharmaceutical composition according to any one of embodiments 62 to 158 and 161, wherein the target cells are pancreatic cells.

[0489] Embodiment 164. A pharmaceutical composition according to any one of embodiments 62 to 158, wherein the target cells are pancreatic cancer cells.

[0490] Embodiment 165. A pharmaceutical composition according to any one of embodiments 62 to 158, wherein the target cells are gastrointestinal cells.

[0491] Embodiment 166. A pharmaceutical composition according to any one of embodiments 62 to 158, wherein the target cells are colorectal cancer cells.

[0492] Embodiment 167. A method for delivering a cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition or synthetic delivery system described in any one of the preceding embodiments.

[0493] Embodiment 168. The method of embodiment 167, wherein the cargo is delivered to an intracellular compartment of a target cell.

[0494] Embodiment 169. The method of embodiment 167 or 168, wherein the cargo is delivered to the nucleus of the target cell.

[0495] Embodiment 170. The method of any one of embodiments 167 to 169, wherein the cargo comprises a nucleic acid cargo, and the nucleic acid cargo is integrated into the genome of the target cell.

[0496] Embodiment 171. The method of embodiment 170, wherein the nucleic acid cargo is integrated into the genome of the target cell at a location identified in Table 2, Table 4, Table 7, or Table 8.

[0497] Embodiment 172. The method of embodiment 170, wherein the nucleic acid cargo is integrated into the genome of the target cell at Chr2:32916224-32916626.

[0498] Embodiment 173. The method of embodiment 170, wherein the nucleic acid cargo is integrated into the genome of the target cell at Ch16:32628381-32629000.

[0499] Embodiment 174. A method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described in any one of the preceding embodiments.

[0500] Embodiment 175. The method of embodiment 174, wherein the subject is a mammal.

[0501] Embodiment 176. The method of any one of embodiments 174, wherein the subject is a human.

[0502] Embodiment 177. The method of any one of embodiments 174 to 176, wherein the condition is cancer.

[0503] Embodiment 178. The method of embodiment 177, wherein the cancer is a hematological cancer.

[0504] Embodiment 179. The method of embodiment 177, wherein the cancer is multiple myeloma.

[0505] Embodiment 180. The method of embodiment 177, wherein the cancer is a solid tumor.

[0506] Embodiment 181. The method of embodiment 177 or 180, wherein the cancer is pancreatic cancer.

[0507] Embodiment 182. The method of embodiment 177 or 180, wherein the cancer is colorectal cancer.

[0508] Embodiment 183. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to any one of SEQ ID NOs: 73-75.

[0509] Embodiment 184. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 73.

[0510] Embodiment 185. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 73.

[0511] Embodiment 186. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 73.

[0512] Embodiment 187. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of SEQ ID NO: 73.

[0513] Embodiment 188. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 74.

[0514] Embodiment 189. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 74.

[0515] Embodiment 190. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 74.

[0516] Embodiment 191. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of SEQ ID NO: 74.

[0517] Embodiment 192. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 75.

[0518] Embodiment 193. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 75.

[0519] Embodiment 194. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 75.

[0520] Embodiment 195. The pharmaceutical composition of embodiment 183, wherein the recognition sequence consists essentially of SEQ ID NO: 75.

[0521] Embodiment 196. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to SEQ ID NO: 71.

[0522] Embodiment 197. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 71.

[0523] Embodiment 198. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 71.

[0524] Embodiment 199. A pharmaceutical composition according to any one of embodiments 183 to 195, wherein the recognition sequence does not contain the nucleotide sequence of SEQ ID NO: 71.

[0525] Embodiment 200. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to nucleotides 1 to 87 of SEQ ID NO: 68.

[0526] Embodiment 201. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to nucleotides 1 to 87 of SEQ ID NO: 68.

[0527] Embodiment 202. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to nucleotides 1 to 87 of SEQ ID NO: 68.

[0528] Embodiment 203. The pharmaceutical composition of any one of embodiments 183 to 195, wherein the recognition sequence does not contain the nucleotide sequence of nucleotides 1 to 87 of SEQ ID NO: 68.

[0529] Embodiment 204. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 73.

[0530] Embodiment 205. The pharmaceutical composition of embodiment 204, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 74.

[0531] Embodiment 206. The pharmaceutical composition of embodiment 204, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 74.

[0532] Embodiment 207. The pharmaceutical composition of embodiment 204, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 74.

[0533] Embodiment 208. The pharmaceutical composition of embodiment 204, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 75.

[0534] Embodiment 209. The pharmaceutical composition of embodiment 204, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 75.

[0535] Embodiment 210. The pharmaceutical composition of embodiment 204, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 75.

[0536] Embodiment 211. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to SEQ ID NO: 71.

[0537] Embodiment 212. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 71.

[0538] Embodiment 213. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 71.

[0539] Embodiment 214. A pharmaceutical composition according to any one of embodiments 204 to 210, wherein the recognition sequence does not contain the nucleotide sequence of SEQ ID NO: 71.

[0540] Embodiment 215. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to nucleotides 1 to 87 of SEQ ID NO: 68.

[0541] Embodiment 216. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to nucleotides 1 to 87 of SEQ ID NO: 68.

[0542] Embodiment 217. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to nucleotides 1 to 87 of SEQ ID NO: 68.

[0543] Embodiment 218. The pharmaceutical composition of any one of embodiments 204 to 210, wherein the recognition sequence does not contain the nucleotide sequence of nucleotides 1 to 87 of SEQ ID NO: 68.

[0544] Embodiment 219. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by a target cell, and wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13 or 14.

[0545] Embodiment 220. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13.

[0546] Embodiment 221. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 13.

[0547] Embodiment 222. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 13.

[0548] Embodiment 223. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 13.

[0549] Embodiment 224. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 13.

[0550] Embodiment 225. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13.

[0551] Embodiment 226. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 13.

[0552] Embodiment 227. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 13.

[0553] Embodiment 228. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 13.

[0554] Embodiment 229. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 13.

[0555] Embodiment 230. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 13.

[0556] Embodiment 231. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 13.

[0557] Embodiment 232. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 13.

[0558] Embodiment 233. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 14.

[0559] Embodiment 234. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 14.

[0560] Embodiment 235. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 14.

[0561] Embodiment 236. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 14.

[0562] Embodiment 237. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 14.

[0563] Embodiment 238. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 14.

[0564] Embodiment 239. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 25 consecutive nucleotides of SEQ ID NO: 14.

[0565] Embodiment 240. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 14.

[0566] Embodiment 241. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 14.

[0567] Embodiment 242. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 14.

[0568] Embodiment 243. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 14.

[0569] Embodiment 244. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 14.

[0570] Embodiment 245. The pharmaceutical composition of embodiment 219, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 14.

[0571] Embodiment 246. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell, and wherein the recognition sequence comprises (a) a nucleotide sequence having at least 90% sequence identity to at least 60 contiguous nucleotides of SEQ ID NO:33, or (b) a nucleotide sequence having at least 91% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:33.

[0572] Embodiment 247. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 60 consecutive nucleotides of SEQ ID NO: 33.

[0573] Embodiment 248. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 91% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 33.

[0574] Embodiment 249. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 75 consecutive nucleotides of SEQ ID NO: 33.

[0575] Embodiment 250. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 consecutive nucleotides of SEQ ID NO: 33.

[0576] Embodiment 251. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 consecutive nucleotides of SEQ ID NO: 33.

[0577] Embodiment 252. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO: 33.

[0578] Embodiment 253. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 33.

[0579] Embodiment 254. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 33.

[0580] Embodiment 255. The pharmaceutical composition of embodiment 246, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO: 33.

[0581] Embodiment 256. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, and the recognition sequence comprises an AluSp, AluSx, AluSg2, ERV2 superfamily or ERV3 superfamily transposable element or a functional fragment thereof that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell.

[0582] Embodiment 257. The pharmaceutical composition of embodiment 256, wherein the recognition sequence comprises an AluSp transposable element or a functional fragment thereof.

[0583] Embodiment 258. The pharmaceutical composition of embodiment 256, wherein the recognition sequence comprises an AluSx transposable element or a functional fragment thereof.

[0584] Embodiment 259. The pharmaceutical composition of embodiment 256, wherein the recognition sequence comprises an AluSg2 transposable element or a functional fragment thereof.

[0585] Embodiment 260. The pharmaceutical composition of embodiment 256, wherein the recognition sequence comprises an ERV2 superfamily transposable element or a functional fragment thereof.

[0586] Embodiment 261. A pharmaceutical composition described in embodiment 260, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises an HML8 group transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0587] Embodiment 262. The pharmaceutical composition of embodiment 260, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises a MER family transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by the target cell.

[0588] Embodiment 263. A pharmaceutical composition described in embodiment 260, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises a MER11C transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0589] Embodiment 264. The pharmaceutical composition of embodiment 256, wherein the recognition sequence comprises an ERV3 superfamily transposable element or a functional fragment thereof.

[0590] Embodiment 265. A pharmaceutical composition described in embodiment 264, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises a MaLR group transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0591] Embodiment 266. A pharmaceutical composition described in embodiment 264, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises a THE1A transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell.

[0592] Embodiment 267. A pharmaceutical composition described in embodiment 264, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises an MLT1J2 transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or its components by the target cell.

[0593] Embodiment 268. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or a component thereof by the target cell, and wherein the polynucleotide comprises a nucleotide sequence having at least 86% sequence identity to any one of SEQ ID NOs: 7, 16, 25, 29 and 31.

[0594] Embodiment 269. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:7.

[0595] Embodiment 270. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 16.

[0596] Embodiment 271. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 25.

[0597] Embodiment 272. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 29.

[0598] Embodiment 273. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 31.

[0599] Embodiment 274. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:7.

[0600] Embodiment 275. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 16.

[0601] Embodiment 276. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 25.

[0602] Embodiment 277. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 29.

[0603] Embodiment 278. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 31.

[0604] Embodiment 279. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:7.

[0605] Embodiment 280. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 16.

[0606] Embodiment 281. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 25.

[0607] Embodiment 282. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 29.

[0608] Embodiment 283. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO: 31.

[0609] Embodiment 284. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 7.

[0610] Embodiment 285. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 16.

[0611] Embodiment 286. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 25.

[0612] Embodiment 287. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 29.

[0613] Embodiment 288. The pharmaceutical composition of embodiment 268, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 31.

[0614] Embodiment 289. A pharmaceutical composition according to any one of embodiments 183 to 288, wherein the recognition sequence facilitates binding of the synthetic delivery system to target cells.

[0615] Embodiment 290. A pharmaceutical composition according to any one of embodiments 183 to 289, wherein the recognition sequence facilitates binding of the synthetic delivery system to target cells that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

[0616] Embodiment 291. A pharmaceutical composition according to any one of embodiments 183 to 290, wherein the recognition sequence facilitates binding of the synthetic delivery system to target cells that is at least 50% higher compared to control cells of a different cell type.

[0617] Embodiment 292. A pharmaceutical composition according to any one of embodiments 183 to 291, wherein the recognition sequence facilitates uptake of the synthetic delivery system or a component thereof by target cells.

[0618] Embodiment 293. A pharmaceutical composition according to any one of embodiments 183 to 292, wherein the recognition sequence facilitates uptake of the synthetic delivery system by target cells that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

[0619] Embodiment 294. A pharmaceutical composition according to any one of embodiments 183 to 293, wherein the recognition sequence facilitates uptake of the synthetic delivery system by target cells that is at least 50% higher compared to control cells of a different cell type.

[0620] Embodiment 295. A pharmaceutical composition described in any one of embodiments 183 to 294, wherein the recognition sequence facilitates nuclear localization of the synthetic delivery system within the target cell that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

[0621] Embodiment 296. A pharmaceutical composition according to any one of embodiments 183 to 295, wherein the recognition sequence facilitates nuclear localization of the synthetic delivery system in target cells that is at least 50% higher compared to control cells of a different cell type.

[0622] Embodiment 297. A pharmaceutical composition according to any one of embodiments 183 to 296, wherein the recognition sequence facilitates binding of the synthetic delivery system to and uptake of the synthetic delivery system or its components by the target cells.

[0623] Embodiment 298. A pharmaceutical composition according to any one of embodiments 183 to 297, wherein the recognition sequence comprises a nucleic acid sequence identified in circulating tumor DNA (ctDNA) from a cell type substantially similar to the target cell.

[0624] Embodiment 299. The pharmaceutical composition of any one of embodiments 183 to 298, wherein the polynucleotide comprises double-stranded DNA.

[0625] Embodiment 300. The pharmaceutical composition of any one of embodiments 183 to 299, wherein the synthetic delivery system further comprises a cargo.

[0626] Embodiment 301. The pharmaceutical composition of embodiment 300, wherein the cargo comprises an anti-cancer therapeutic agent.

[0627] Embodiment 302. A pharmaceutical composition according to embodiment 300 or 301, wherein the cargo comprises a nucleic acid cargo.

[0628] Embodiment 303. The pharmaceutical composition of embodiment 302, wherein the nucleic acid cargo encodes a tumor suppressor protein.

[0629] Embodiment 304. A pharmaceutical composition according to any one of embodiments 300 to 303, wherein the cargo is attached to the 3' end of the polynucleotide.

[0630] Embodiment 305. A pharmaceutical composition according to any one of embodiments 300 to 304, wherein the cargo is attached to the 5' end of the polynucleotide.

[0631] Embodiment 306. A pharmaceutical composition according to any one of embodiments 300 to 305, wherein the cargo comprises a cytotoxic cargo.

[0632] Embodiment 307. A pharmaceutical composition according to any one of embodiments 300 to 306, wherein the cargo comprises a therapeutic cargo.

[0633] Embodiment 308. A pharmaceutical composition according to any one of embodiments 300 to 307, wherein the cargo comprises a polypeptide.

[0634] Embodiment 309. A pharmaceutical composition according to any one of embodiments 300 to 308, wherein the cargo comprises a small molecule.

[0635] Embodiment 310. The pharmaceutical composition of any one of embodiments 183 to 309, wherein the synthetic delivery system further comprises a promoter.

[0636] Embodiment 311. The pharmaceutical composition of any one of embodiments 183 to 310, wherein the synthetic delivery system further comprises an integration sequence that facilitates integration of the polynucleotide, synthetic delivery system, cargo or a component thereof into the genome of the target cell.

[0637] Embodiment 312. The pharmaceutical composition of embodiment 311, wherein the integration sequence facilitates genomic integration of the synthetic delivery system, polynucleotide, cargo or component thereof in the target cell that is at least 50% higher compared to a control synthetic delivery system lacking the integration sequence.

[0638] Embodiment 313. A pharmaceutical composition according to any one of embodiments 311 to 312, wherein the integration sequence facilitates genomic integration of the synthetic delivery system, polynucleotide, cargo or a component thereof by the target cell, which integration sequence is at least 50% higher compared to control cells of a different cell type.

[0639] Embodiment 314. A pharmaceutical composition described in any one of embodiments 183 to 313, wherein the synthetic delivery system does not utilize a viral vector, nanoparticle, lipid nanoparticle, liposome, exosome, dendrimer, gene gun or electroporation for delivery to target cells.

[0640] Embodiment 315. A pharmaceutical composition according to any one of embodiments 183 to 314, wherein the target cells are leukocytes.

[0641] Embodiment 316. A pharmaceutical composition according to any one of embodiments 183 to 315, wherein the target cells are plasma cells.

[0642] Embodiment 317. A pharmaceutical composition described in any one of embodiments 183 to 316, wherein the target cells are cancer cells.

[0643] Embodiment 318. A pharmaceutical composition according to any one of embodiments 183 to 317, wherein the target cells are multiple myeloma cells.

[0644] Embodiment 319. A pharmaceutical composition according to any one of embodiments 183 to 314, wherein the target cells are pancreatic cells.

[0645] Embodiment 320. A pharmaceutical composition described in any one of embodiments 183 to 314, wherein the target cells are pancreatic cancer cells.

[0646] Embodiment 321. A pharmaceutical composition according to any one of embodiments 183 to 314, wherein the target cells are gastrointestinal cells.

[0647] Embodiment 322. A pharmaceutical composition according to any one of embodiments 183 to 314, wherein the target cells are colorectal cancer cells.

[0648] Embodiment 323. A method for delivering a cargo to a target cell, comprising contacting the target cell with a pharmaceutical composition or synthetic delivery system described in any one of the preceding embodiments.

[0649] Embodiment 324. The method of embodiment 323, wherein the cargo is delivered to an intracellular compartment of a target cell.

[0650] Embodiment 325. The method of any one of embodiments 323-324, wherein the cargo is delivered to the nucleus of the target cell.

[0651] Embodiment 326. The method of any one of embodiments 323 to 325, wherein the cargo comprises a nucleic acid cargo, and the nucleic acid cargo is integrated into the genome of the target cell.

[0652] Embodiment 327 The method of any one of embodiments 323 to 326, wherein the nucleic acid cargo is integrated into the genome of the target cell at a location identified in Table 2, Table 4, Table 7 or Table 8.

[0653] Embodiment 328. The method of any one of embodiments 323 to 327, wherein the nucleic acid cargo is integrated into the genome of the target cell at Chr2:32916224-32916626.

[0654] Embodiment 329. The method of any one of embodiments 323 to 327, wherein the nucleic acid cargo is integrated into the genome of the target cell at Ch16:32628381-32629000.

[0655] Embodiment 330. A method of treating a condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described in any one of the preceding embodiments.

[0656] Embodiment 331. The method of embodiment 330, wherein the subject is a mammal.

[0657] Embodiment 332. The method of any one of embodiments 330 to 331, wherein the subject is a human.

[0658] Embodiment 333. The method of any one of embodiments 330 to 332, wherein the condition is cancer.

[0659] Embodiment 334. The method of embodiment 333, wherein the cancer is a hematological cancer.

[0660] Embodiment 335. The method of any one of embodiments 333 to 334, wherein the cancer is multiple myeloma.

[0661] Embodiment 336. The method of embodiment 333, wherein the cancer is a solid tumor.

[0662] Embodiment 337. The method of embodiment 333, wherein the cancer is pancreatic cancer.

[0663] Embodiment 338. The method of any one of embodiments 333, wherein the cancer is colorectal cancer. [Example]

[0664] VII. Working Examples Example 1 Identification of genomic integration sites of ctDNA We assessed the integration of ctDNA into the host cell genome. ctDNA was extracted from patient plasma and cultured with matched or mismatched cancer cell lines for 24 hours, followed by whole-genome sequencing. The cell lines used were MM1s (multiple myeloma) and MIA (pancreatic cancer).

[0665] We identified single-nucleotide variants (SNVs) unique to ctDNA that were found in the host cell genome under matched coculture conditions. Comparison of sequences from each experimental condition after alignment with the reference human genome (Hg38) led to the identification of multiple SNVs shared between the coculture conditions and ctDNA that were not present in the genome of untreated control cells (referred to herein as "SNVs of interest," Figure 1A). Further elaboration of these SNVs of interest demonstrated that cells cocultured with ctDNA exhibited a skew in variant allele frequency relative to that of the variant ctDNA allele, compared to cells alone (Figure 1B). Examples of SNVs demonstrating ctDNA "skew" in variant allele frequency (VAF) after coculture with ctDNA are shown in Figures 2A-2D.

[0666] Using the sequence data, we reconstructed genomic contigs for each experimental condition by de novo assembly. Comparative analysis of contig sequences between the cell lines, ctDNA, and coculture conditions described above resulted in the detection of ctDNA fragments integrated into the cellular genome under coculture conditions. NucDiff analysis was used to identify coculture contigs harboring ctDNA insertions. BLAST alignment analysis further verified the acquisition of ctDNA fragments in the coculture conditions and identified the transition points of ctDNA insertion (Figures 3A-C). Analysis of the integrated ctDNA fragments and genomic insertion sites identified from both tumor models indicated that the majority (nearly 67%) of the inserted ctDNA fragments originated from chromosomes 3 and 7. Furthermore, approximately 80% of the ctDNA fragments targeted cellular chromosomal regions near the genomic location of the ctDNA origin, suggesting a high potential for ctDNA insertion in homologous target regions. The remaining 20% inserted into chromosomal locations distinct from their site of origin. Example 2 Identification of transposable elements in ctDNA

[0667] Experiments were designed to identify the presence of transposons in ctDNA fragments. To more precisely map ctDNA genomic junctions, PACBIO probes were ligated to multiple myeloma and pancreatic cancer ctDNA samples to label their 5' and 3' ends. Whole-genome sequencing of DNA from cells with ctDNA genomic integration was performed using the Illumina platform. De novo assembly was performed on the sequencing output. ctDNA contigs containing PACBIO adapters were then matched to contigs containing inserted ctDNA sequences identified in coculture experiments. Transposable elements (<100 nucleotides from the transition point between the cell and ctDNA) on the ctDNA sequences at the insertion point were detected and classified using RepeatMaster. This analysis demonstrated that ctDNA fragments integrated into the cellular genome contained more transposable elements than unintegrated ctDNA fragments (Figure 4). These findings suggested that retrotransposons mediate horizontal gene transfer in cancer cells.

[0668] We searched for transposons that preferentially target cells of the matching tumor type. We compared the lists of transposons identified at ctDNA insertion points in matched and mismatched co-culture conditions (e.g., MM ctDNA with MM cells or PC cells). We selected transposons that were uniquely or preferentially inserted in the matched condition.

[0669] Class I transposons ERV-L, LTR, and SINE and LINE comprised the majority of transposable elements at transition points in pancreatic cancer or multiple myeloma.

[0670] Within the matched PC ctDNA, AluSx, MIRc, and MTL1J were from the most common retrotransposon subfamilies located at the insertion points (Fig. 5A and Table 8).

[0671] In multiple myeloma, AluSp, MER11, MER11C, AluJb, and L2a, among other subfamilies of retrotransposons, were identified at MM ctDNA insertion sites (Figure 5B and Table 7).

[0672] Activation of short interspersed nucleotide sequences (SINEs) and long interspersed nucleotide sequences (LINEs) was observed in cancer, leaving the possibility of retrotransposition. Expression of transposable elements was assessed in RNA sequencing data from 60 MM and 23 PC patient samples. Batch-normalized raw data was used to measure expression variability. Analysis identified several tissue-specific retrotransposons highly expressed across pancreatic cancer and multiple myeloma. These findings demonstrate that the retrotransposons identified at insertion sites are transcriptionally active in these cancer types (Figures 6A and 6B).

[0673] To functionally assess the role of retrotransposons in horizontal gene transfer of ctDNA, experiments were performed to examine the effects of reverse transcriptase or integrase inhibition on ctDNA integration. Multiple myeloma (MM1s), pancreatic cancer (MIA), and colon cancer (HCT116) cell lines were treated with the reverse transcriptase inhibitors zidovudine or didanosine and the integrase inhibitor raltegravir before adding ctDNA to the culture medium. Treatment with reverse transcriptase or integrase inhibitors significantly reduced ctDNA chromatid integration compared to untreated controls (Figure 7), providing evidence that retrotransposons play an important role in horizontal gene transfer of ctDNA. Example 3 Evaluation of synthesized transposable elements and cargo delivery

[0674] Several MM retrotransposons containing tumor-type-specific point mutations were synthesized and tested for their ability to integrate. Retrotransposons that could be synthesized using the gBlock gene fragment method were selected (including AluSp, AluSg2, MER11C, THE1A, and AluSx). For controls, a PC retrotransposon (AluSx1) and a synthetic sequence of similar length and GC content that did not contain retrotransposon sequences were included.

[0675] The ability of the retrotransposons to resist degradation induced by DNAse contained in the complete culture medium was assessed. Agarose gel electrophoresis demonstrated that all fragments remained intact under the culture conditions (Figure 8).

[0676] Retrotransposons were labeled with CY5 to determine uptake by MM cell lines using flow cytometry. The kinetics of retrotransposon capture were monitored by flow cytometry. Results showed that cellular capture of CY5-AluSp increased over time more efficiently than the control sequence (Figure 9A).

[0677] A dose-finding experiment was performed to assess cell capture and internalization. DNA was considered internalized if it was not removed from the cells by trypsin treatment. This experiment demonstrated dose-dependent CY5-AluSp cell capture and internalization that was more efficient than that observed with the control sequence (Figure 9B).

[0678] The variability of cell capture of several transposons was examined in two MM cell lines at 4 h of culture. These experiments determined that cell capture of CY5-AluSp and CY5-MER11C was higher than that of other transposons, control sequences, and the PC transposon (Figures 9C and 9D).

[0679] The cell targeting of AluSp and MER11C sequences was examined in bone marrow samples from patients with and without multiple myeloma. After 14 hours of cell culture, more CY5-positive plasma cells were observed in samples cultured with CY5-AluSp or CY5-MER11C (Figure 10, upper panel) than in samples cultured with the control or PC-specific sequences (Figure 10, lower left panel). Interestingly, the level of plasma cell targeting was higher in plasma cells from MM patients than in those without MM. This result suggests that malignant plasma cells are more suitable for capturing these retrotransposons (Figure 11). AluSp and MER11C showed higher levels of signal in plasma cells from MM patients than in non-MM CD138(-) cell compartments, verifying their tissue-specific cell targeting ability (Figure 10, lower right panel, and Figure 11).

[0680] To verify that the retrotransposon sequence contains a cell recognition signal, we generated several deletion mutants of the AluSp sequence and evaluated the effect of the deletions on cell capture. The results showed that the absence of the 3' end of the AluSp sequence reduced MM cell capture, suggesting that the last approximately 230 base pairs contain the MM cell recognition sequence (Figures 12A and 12B). Multiple sequence alignment of AluSp and all MM-specific transposons revealed several conserved nucleotides and two GC- and AT-rich regions toward the 3' end (Figure 12C).

[0681] Experiments were designed to examine the ability of AluSp sequences to transfer genetic material between cells. We examined the delivery of an mCherry reporter cassette into MM cell lines using AluSp sequences. To do this, AluSp was ligated to both ends of a linearized CMV promoter-mCherry cassette. This AluSp-CMV-mCherry ligation product was then added to the culture medium of MM1s cells for 24 hours, and mCherry expression was assessed. Microscopy experiments demonstrated a higher number of mCherry-expressing cells in AluSp-CMV-mCherry-treated cells compared to cells treated with the linearized vector alone or after transfection with the circularized CMV-mCherry vector (Figure 13). Expression of mCherry in AluSp-CMV-mCherry-mediated MM1s cell cultures was also detected by flow cytometry.

[0682] To assess genomic integration of the AluSp-CMV-mCherry cassette, whole-genome single-cell sequencing was performed in MM1s cells expressing various levels of mCherry. mCherry insertions were identified by recognizing sequences in which one read aligned to the cellular genome and the other aligned to either the CMV or mCherry sequence. This analysis detected various levels of mCherry insertion in cells with moderate or high expression levels, while no insertion was identified in cells that failed to express mCherry (Figure 14A). In contrast, when cells were cultured with the control-mCherry sequence, very few cells were found to have high levels of mCherry insertion and expression. mCherry integration was confirmed by mCherry PCR in cells cultured with the AluSp-mCherry vector, but not in control-mCherry-treated cells (Figure 14B). The AluSp-CMV-mCherry vector integrated with high fidelity into two specific regions in the genome (Chr2:32916224-32916626 and Ch16:32628381-32629000), which are enriched for simple repeats and AluSp, respectively (Figure 14C).

[0683] We assessed AluSp's ability to target tissue-specific cells and deliver payloads by ligating the herpes simplex virus tyrosine kinase (HSV-Tk) cytotoxic gene between the AluSp sequences (AluSp-HSV-Tk-GFP) and examining the effect of the construct on the viability of MM, PC, and CC cell lines. Cells were cultured with AluSp-HSV-Tk-GFP for 24 hours, followed by 96 hours of treatment with ganciclovir (GCV), which is converted to a toxic product by HSV-Tk. GCV specifically reduced cell viability of MM cells treated with MM-AluSp-HSV-Tk (Figure 15). In contrast, AluSp-HSV-Tk / GCV treatment caused no or substantial changes in viability in PC or CC cell lines.

[0684] These results demonstrated that the synthesized AluSp can deliver and incorporate genetic cargo material into specific cells and can be combined with cytotoxic cargo to target cancer cells for killing. Example 4 ctDNA alters the drug response phenotype of MM or PC cell lines

[0685] The effect of ctDNA horizontal gene transfer on cellular drug sensitivity was evaluated.

[0686] Gemcitabine (GEM)-sensitive pancreatic cancer cell lines (MIA Paca-2 [MIA] and PANC1) were cultured with DNase-treated or untreated plasma from GEM-resistant (GR) patients or control plasma for 24 hours. Compared with control serum, both cell lines became resistant to GEM when cultured with non-DNase-treated GR plasma. Exposure to DNase-treated GR plasma restored the cells' inherent sensitivity to treatment (Figure 16). In contrast, treatment of control serum with DNase resulted in increased resistance of PC cells to gemcitabine.

[0687] To further validate the contribution of ctDNA to drug response propagation, cell-free ctDNA was extracted from bortezomib-resistant (BR) or -sensitive (BS) patients and added to culture medium containing plasma from patients without cancer or bortezomib resistance. The results showed that adding ctDNA from BR patients to non-cancer patient plasma reduced the sensitivity of MM1s and OPM1 to bortezomib (Figure 17). Additionally, when BR patient ctDNA was added to BR patient plasma, resistance to bortezomib was further increased. In contrast, adding BS patient ctDNA to RPMI cells cultured with non-cancer patient control plasma resulted in increased sensitivity to bortezomib. These findings suggest that ctDNA can transmit genetic material that confers drug sensitivity or resistance from one cell to another. Example 5 Additional Materials and Methods

[0688] Clinical Specimens and Sample Preparation: Retrospective plasma samples were obtained from 10 patients with multiple myeloma (MM), 10 with pancreatic cancer (PC), 3 with colon cancer (CC), and 2 with lung cancer (LC) from samples stored at Tissue and Acquisition. Ten patients with newly diagnosed MM were treated with bortezomib-containing regimens (five responders and five non-responders). Seven of the 10 patients with PC had advanced-stage disease and were treated with gemcitabine (two patients had a partial response and five had progressive disease at the time of plasma sample acquisition), and three early-stage patients underwent surgical resection. Responses in MM were determined using the International Uniform Response Criteria for Multiple Myeloma (UNIM), and in PC patients, responses were evaluated using the Response Evaluation Criteria in Solid Tumors (RECIST) criteria. The blood samples were centrifuged at 1500 rpm for 10 minutes, after which plasma was isolated. The plasma supernatant was collected for storage. Plasma from non-cancer patients was purchased from a commercial vendor.

[0689] Cell lines and reagents: The following cell lines were grown in Roswell Park Memorial Institute (RPMI)-1640 medium: four multiple myeloma (OPM1, RPMI, JK6L, and MM1S); one PC (ASPC-1); and one LC (A549). Col...

Claims

1. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell, and wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to any one of SEQ ID NOs:73-75.

2. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

73.

3. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

73.

4. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

73.

5. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of SEQ ID NO:

73.

6. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

74.

7. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

74.

8. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

74.

9. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of SEQ ID NO:

74.

10. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

75.

11. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

75.

12. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

75.

13. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence consists essentially of the nucleotide sequence of SEQ ID NO:

75.

14. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to SEQ ID NO:

71.

15. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

71.

16. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

71.

17. The pharmaceutical composition of claim 1 , wherein the recognition sequence does not contain the nucleotide sequence of SEQ ID NO:

71.

18. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to nucleotides 1-87 of SEQ ID NO:

68.

19. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to nucleotides 1-87 of SEQ ID NO:

68.

20. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to nucleotides 1-87 of SEQ ID NO:

68.

21. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence does not contain the nucleotide sequence of nucleotides 1 to 87 of SEQ ID NO:

68.

22. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell, and the recognition sequence comprises the nucleotide sequence of SEQ ID NO:

73.

23. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

74.

24. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

74.

25. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO:

74.

26. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

75.

27. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

75.

28. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO:

75.

29. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to SEQ ID NO:

71.

30. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to SEQ ID NO:

71.

31. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

71.

32. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain the nucleotide sequence of SEQ ID NO:

71.

33. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain a nucleotide sequence having at least 50% sequence identity to nucleotides 1-87 of SEQ ID NO:

68.

34. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain a nucleotide sequence having at least 80% sequence identity to nucleotides 1-87 of SEQ ID NO:

68.

35. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain a nucleotide sequence having at least 90% sequence identity to nucleotides 1-87 of SEQ ID NO:

68.

36. 23. The pharmaceutical composition of claim 22, wherein the recognition sequence does not contain the nucleotide sequence of nucleotides 1 to 87 of SEQ ID NO:

68.

37. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell, and the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO: 13 or 14.

38. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO:

13.

39. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 25 contiguous nucleotides of SEQ ID NO:

13.

40. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:

13.

41. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO:

13.

42. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 200 contiguous nucleotides of SEQ ID NO:

13.

43. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO:

13.

44. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 25 contiguous nucleotides of SEQ ID NO:

13.

45. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:

13.

46. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO:

13.

47. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 contiguous nucleotides of SEQ ID NO:

13.

48. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

13.

49. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

13.

50. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO:

13.

51. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO:

14.

52. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 25 contiguous nucleotides of SEQ ID NO:

14.

53. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:

14.

54. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO:

14.

55. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 80% sequence identity to at least 200 contiguous nucleotides of SEQ ID NO:

14.

56. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 10 consecutive nucleotides of SEQ ID NO:

14.

57. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 25 contiguous nucleotides of SEQ ID NO:

14.

58. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:

14.

59. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO:

14.

60. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 contiguous nucleotides of SEQ ID NO:

14.

61. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

14.

62. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

14.

63. 38. The pharmaceutical composition of claim 37, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO:

14.

64. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or a component thereof by the target cell, and the recognition sequence comprises (a) a nucleotide sequence having at least 90% sequence identity to at least 60 consecutive nucleotides of SEQ ID NO:33, or (b) a nucleotide sequence having at least 91% sequence identity to at least 50 consecutive nucleotides of SEQ ID NO:

33.

65. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 60 contiguous nucleotides of SEQ ID NO:

33.

66. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 91% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:

33.

67. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 75 contiguous nucleotides of SEQ ID NO:

33.

68. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 100 contiguous nucleotides of SEQ ID NO:

33.

69. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to at least 200 contiguous nucleotides of SEQ ID NO:

33.

70. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to at least 50 contiguous nucleotides of SEQ ID NO:

33.

71. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

33.

72. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

33.

73. 65. The pharmaceutical composition of claim 64, wherein the recognition sequence comprises the nucleotide sequence of SEQ ID NO:

33.

74. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence, the recognition sequence comprising an AluSp, AluSx, AluSg2, ERV2 superfamily or ERV3 superfamily transposable element or a functional fragment thereof that facilitates binding of the synthetic delivery system to or uptake of the synthetic delivery system or a component thereof by a target cell.

75. 75. The pharmaceutical composition of claim 74, wherein the recognition sequence comprises the AluSp transposable element or a functional fragment thereof.

76. 75. The pharmaceutical composition of claim 74, wherein the recognition sequence comprises the AluSx transposable element or a functional fragment thereof.

77. 75. The pharmaceutical composition of claim 74, wherein the recognition sequence comprises the AluSg2 transposable element or a functional fragment thereof.

78. 75. The pharmaceutical composition of claim 74, wherein the recognition sequence comprises the ERV2 superfamily transposable element or the functional fragment thereof.

79. The pharmaceutical composition of claim 78, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises an HML8 group transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to the target cell or uptake of the synthetic delivery system or its components by the target cell.

80. The pharmaceutical composition of claim 78, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises a MER family transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to the target cell or uptake of the synthetic delivery system or its components by the target cell.

81. The pharmaceutical composition of claim 78, wherein the ERV2 superfamily transposable element or functional fragment thereof comprises a MER11C transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to the target cell or uptake of the synthetic delivery system or its components by the target cell.

82. 75. The pharmaceutical composition of claim 74, wherein the recognition sequence comprises the ERV3 superfamily transposable element or the functional fragment thereof.

83. The pharmaceutical composition of claim 82, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises a MaLR group transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to the target cell or uptake of the synthetic delivery system or its components by the target cell.

84. The pharmaceutical composition of claim 82, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises a THE1A transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to the target cell or uptake of the synthetic delivery system or its components by the target cell.

85. The pharmaceutical composition of claim 82, wherein the ERV3 superfamily transposable element or functional fragment thereof comprises an MLT1J2 transposable element or functional fragment thereof that facilitates binding of the synthetic delivery system to the target cell or uptake of the synthetic delivery system or its components by the target cell.

86. A pharmaceutical composition comprising a synthetic delivery system and a pharmaceutically acceptable excipient, wherein the synthetic delivery system comprises a polynucleotide comprising a recognition sequence that facilitates binding of the synthetic delivery system to a target cell or uptake of the synthetic delivery system or its components by the target cell, and the polynucleotide comprises a nucleotide sequence having at least 86% sequence identity to any one of SEQ ID NOs: 7, 16, 25, 29 and 31.

87. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

7.

88. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

16.

89. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

25.

90. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

29.

91. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:

31.

92. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

7.

93. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

16.

94. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

25.

95. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

29.

96. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:

31.

97. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:

7.

98. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:

16.

99. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:

25.

100. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:

29.

101. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises a nucleotide sequence having at least 98% sequence identity to SEQ ID NO:

31.

102. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:

7.

103. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:

16.

104. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:

25.

105. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:

29.

106. 87. The pharmaceutical composition of claim 86, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:

31.

107. 2. The pharmaceutical composition of claim 1, wherein the polynucleotide comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO:

68.

108. The pharmaceutical composition of claim 1 , wherein the polynucleotide comprises the amino acid sequence of SEQ ID NO:

68.

109. The pharmaceutical composition of claim 1 , wherein the recognition sequence facilitates binding of the synthetic delivery system to the target cell.

110. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates at least 50% greater binding of the synthetic delivery system to the target cell compared to a control synthetic delivery system lacking the recognition sequence.

111. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates at least 50% greater binding of the synthetic delivery system to the target cells compared to control cells of a different cell type.

112. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates uptake of the synthetic delivery system or a component thereof by the target cell.

113. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates at least 50% greater uptake of the synthetic delivery system by the target cells compared to a control synthetic delivery system lacking the recognition sequence.

114. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates uptake of the synthetic delivery system by the target cells that is at least 50% greater than control cells of a different cell type.

115. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates nuclear localization of the synthetic delivery system within the target cell that is at least 50% greater than a control synthetic delivery system lacking the recognition sequence.

116. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates nuclear localization of the synthetic delivery system in the target cell that is at least 50% greater than in a control cell of a different cell type.

117. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence facilitates binding of the synthetic delivery system to the target cell and uptake of the synthetic delivery system or a component thereof by the target cell.

118. 2. The pharmaceutical composition of claim 1, wherein the recognition sequence comprises a nucleic acid sequence identified in circulating tumor DNA (ctDNA) from a cell type substantially similar to the target cell.

119. The pharmaceutical composition of claim 1 , wherein the polynucleotide comprises double-stranded DNA.

120. The pharmaceutical composition of claim 1 , wherein the synthetic delivery system further comprises a cargo.

121. 121. The pharmaceutical composition of claim 120, wherein the cargo comprises an anti-cancer therapeutic agent.

122. 121. The pharmaceutical composition of claim 120, wherein the cargo comprises a nucleic acid cargo.

123. 123. The pharmaceutical composition of claim 122, wherein the nucleic acid cargo encodes a tumor suppressor protein.

124. 121. The pharmaceutical composition of claim 120, wherein the cargo is attached to the 3' end of the polynucleotide.

125. 121. The pharmaceutical composition of claim 120, wherein the cargo is attached to the 5' end of the polynucleotide.

126. 121. The pharmaceutical composition of claim 120, wherein the cargo comprises a cytotoxic cargo.

127. 121. The pharmaceutical composition of claim 120, wherein the cargo comprises a therapeutic cargo.

128. 121. The pharmaceutical composition of claim 120, wherein the cargo comprises a polypeptide.

129. 121. The pharmaceutical composition of claim 120, wherein the cargo comprises a small molecule.

130. 121. The pharmaceutical composition of claim 120, wherein the synthetic delivery system further comprises a promoter.

131. 2. The pharmaceutical composition of claim 1, wherein the synthetic delivery system further comprises an integration sequence that facilitates integration of the polynucleotide, synthetic delivery system, cargo or a component thereof into the genome of the target cell.

132. The pharmaceutical composition of claim 131, wherein the integration sequence facilitates genomic integration of the synthetic delivery system, polynucleotide, cargo or component thereof in the target cell that is at least 50% greater than a control synthetic delivery system lacking the integration sequence.

133. The pharmaceutical composition of claim 131, wherein the integration sequence facilitates genomic integration of the synthetic delivery system, polynucleotide, cargo or component thereof by the target cell that is at least 50% greater than that of a control cell of a different cell type.

134. 10. The pharmaceutical composition of claim 1, wherein the synthetic delivery system does not utilize a viral vector, a nanoparticle, a lipid nanoparticle, a liposome, an exosome, a dendrimer, a gene gun, or electroporation for delivery to the target cells.

135. The pharmaceutical composition of claim 1 , wherein the target cells are leukocytes.

136. The pharmaceutical composition of claim 1 , wherein the target cells are plasma cells.

137. The pharmaceutical composition of claim 1 , wherein the target cell is a cancer cell.

138. The pharmaceutical composition of claim 1 , wherein the target cells are multiple myeloma cells.

139. The pharmaceutical composition of claim 1 , wherein the target cells are pancreatic cells.

140. The pharmaceutical composition of claim 1 , wherein the target cells are pancreatic cancer cells.

141. The pharmaceutical composition of claim 1 , wherein the target cell is a gastrointestinal cell.

142. The pharmaceutical composition of claim 1 , wherein the target cells are colorectal cancer cells.

143. 10. A method of delivering a cargo to a target cell, comprising contacting said target cell with a pharmaceutical composition or synthetic delivery system according to any one of the preceding claims.

144. 144. The method of claim 143, wherein the cargo is delivered to an intracellular compartment of the target cell.

145. 144. The method of claim 143, wherein the cargo is delivered to the nucleus of the target cell.

146. 144. The method of claim 143, wherein the cargo comprises a nucleic acid cargo, and the nucleic acid cargo is integrated into the genome of the target cell.

147. 147. The method of claim 146, wherein said nucleic acid cargo is integrated into the genome of said target cell at a location identified in Table 2, Table 4, Table 7 or Table 8.

148. 147. The method of claim 146, wherein the nucleic acid cargo is integrated into the genome of the target cell at Chr2:32916224-32916626.

149. 147. The method of claim 146, wherein the nucleic acid cargo is integrated into the genome of the target cell at Ch16:32628381-32629000.

150. 10. A method of treating a condition in a subject in need thereof, comprising administering to said subject a pharmaceutical composition according to any one of the preceding claims.

151. 151. The method of claim 150, wherein the subject is a mammal.

152. 151. The method of claim 150, wherein the subject is a human.

153. 151. The method of claim 150, wherein the condition is cancer.

154. 154. The method of claim 153, wherein the cancer is a hematological cancer.

155. 154. The method of claim 153, wherein the cancer is multiple myeloma.

156. 154. The method of claim 153, wherein the cancer is a solid tumor.

157. 154. The method of claim 153, wherein the cancer is pancreatic cancer.

158. 154. The method of claim 153, wherein the cancer is colorectal cancer.

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