46 results about "Retrovirus" patented technology

Among other things, provided herein are systems that replace the natural random integration activity of a retrovirus with site-specific integration machinery. This approach allows for a more precise targeting of a gene of interest into a human genome, e.g., for therapeutic purposes. The system may include integration-deficient retrovirus (e.g., lentivirus) (IDLV), in which the natural integration activity has been reduced (e.g., by mutation to the viral integrase polypeptide). Instead, the system may comprise a site-specific recombinase (e.g., a serine recombinase, e.g., a serine integrase) capable of directing insertion of a template DNA, or portion thereof, into a desired site in the human genome.

Provided herein are immortalized mesenchymal stem cells comprising a replicating recombinant retrovirus. Methods for treating a cell proliferative disorder using these immortalized mesenchymal stem cells are also provided.

The present disclosure belongs to the field of molecular biology, particularly to the technical field of gene analysis and detection, and specifically relates to a method for preparing a DNA library and a method for detecting a retroviral integration site. Specifically, the method for preparing the DNA library comprises the following steps: 1) fragmenting a genomic DNA from a retrovirus-infected cell to obtain DNA fragments; 2) subjecting the DNA fragments to end-repair, A-tailing, and ligation with an adapter to obtain a ligation product, wherein the adapter is an asymmetric double-strand adapter comprising a long-strand sequence and a short-strand sequence, wherein the long-strand sequence sequentially comprises, from a 5' end to a 3' end, a fixed sequence, a random UMI sequence, and an amplification primer binding sequence, and the short-strand sequence comprises a sequence complementary to the fixed sequence; and other steps. The detection method of the present disclosure has extremely high sensitivity, thus having good application potential.

A vaccine for use in the prophylaxis and / or treatment of a diseaseThe present invention relates to an adenoviral vector capable of encoding a virus-like particle (VLP), said VLP displaying an inactive immune-suppressive domain (ISD). The vaccine of the invention shows an improved immune response from either of both of the response pathways initiated by CD4 T cells or CD8 T cells.

The present invention relates to a platform concept for presenting antigenic polypeptides as part of a virus like particle (VLP) construct, which comprises a Gag (group-specific antigen) protein of a beta-retrovirus, for instance of a human endogenous retrovirus K (HERV-K) or of IAPE. Surprisingly it was found that antigenic polypeptide expression in a VLP comprising a Gag protein of HERV-K or of murine endogenous retrovirus IAPE (Intracisternal A-type Particles elements with an Envelope) promotes antigenic polypeptide display and immunogenicity.

The present disclosure provides DNA molecules, such as transfer plasmids, useful for manufacturing packaged retroviral vectors encoding a therapeutic protein such as chimeric antigen receptors (CARs) and T cell receptors (TCRs). The encoded retroviral transcript in the DNA molecule includes a transgene, an elongation factor 1-alpha 1 (EFla) promoter, without the EFla intron, operably linked to the transgene and a Rev response element (RRE).

The present disclosure relates to methods for determining the magnitude of a subject's immune response against a HIVACAT T-cell immunogen (HTI or “HTI immunogen”) and whether the subject can avoid antiretroviral therapy (ART). These methods are helpful for treating human immunodeficiency virus (HIV) and / or deciding whether to administer, continue or stop antiretroviral therapy in a subject. The present disclosure also relates to antigens, compositions, and kits related to such methods.

The invention belongs to the technical field of biological medicines, and discloses an oligonucleotide molecule for targeted inhibition of a human LINC00921 gene, a coding oligopeptide and application of the oligonucleotide molecule in treatment of lung adenocarcinoma. The invention also discloses application of related drugs or products taking the oligonucleotide molecule for targeted inhibition of the human LINC00921 gene as an active ingredient in inhibition or treatment of lung adenocarcinoma. A specific siRNA target sequence is designed aiming at a target gene, a retrovirus vector plasmid containing the target sequence is successfully constructed through genetic engineering, a recombinant retrovirus capable of efficiently silencing the LINC00921 gene is obtained, the mRNA expression level of the LINC00921 gene in lung adenocarcinoma cells can be remarkably reduced, the gene knock-down effect is outstanding, and the retrovirus can be applied to lung adenocarcinoma cells. The compound has a remarkable inhibition effect on proliferation, migration and invasion ability of lung adenocarcinoma cells. Therefore, the invention has important application value in treatment of lung adenocarcinoma, and has high-throughput operability and good repeatability.

The invention provides application of overexpressed CDCA5 in preparation of a medicine for enhancing the anti-tumor function of T cells, and relates to the technical field of cellular immunotherapy. According to the method for overexpressing the CDCA5, a CDCA5 overexpression agent is used, the CDCA5 overexpression agent is a carrier for overexpressing the CDCA5, and the carrier for overexpressing the CDCA5 is any one of lentivirus and retrovirus. The invention overcomes the defects of the prior art, verifies that the CAR-T overexpressing CDCA5 obviously enhances the anti-tumor function in vivo and in vitro, and provides a new technical direction for anti-tumor treatment.

The application belongs to the technical field of biological medicine, and discloses application of a MYLK3 gene inhibitor in preparation of a drug for treating castration-resistant prostate cancer. The biological function of MYLK3 in prostate cancer is confirmed for the first time, an intervention means with the gene as a target point is provided, and it is confirmed that the MYLK3 inhibitor can improve the sensitivity of castration-resistant prostate cancer to androgen receptor antagonists. A gene therapy drug with the human MYLK3 gene as a treatment target point is specially provided, which is suitable for the treatment of prostate cancer (especially castration-resistant prostate cancer), and can also be combined with other targeted drugs to improve the treatment effect. By designing a reasonable RNAi target sequence for the target gene, a retrovirus vector plasmid containing the target sequence is successfully constructed, which has a significant inhibitory effect on the proliferation ability of prostate cancer cells, and significantly increases the sensitivity of prostate cancer cells to AR antagonists.

The present disclosure relates to a kit and a method for purifying retrovirus, which comprises a reagent combination for purifying retrovirus, comprising a first eluent, a second eluent, an elution liquid and a polyanion compound. By using the purification chromatography combination and method of the present disclosure, the viral load can be increased, the chromatography recovery rate of retrovirus is increased by 2-3 times, reaching 80% and above; at the same time, the impurity level in the virus elution liquid is significantly reduced, such as the residual amount of HCP, which can be reduced by 2-3 times; the eluted retrovirus elution liquid can be stored for 24 hours without losing the infection activity under high salt conditions without adding other special protection reagents; after elution, the next step of ultrafiltration dialysis can be directly carried out without the step of dilution and then concentration, effectively simplifying the operation steps and shortening the processing time.

The present disclosure relates to methods of treating or preventing RNA viral infections and retroviral diseases such as HIV and AIDS comprising administering to a subject in need thereof an effective amount of (a) a capsid inhibitor and (b) an adenosine derivative disclosed herein. Also provided are compositions comprising an effective amount of an adenosine derivative and an effective amount of a capsid (CA) inhibitor.

Disclosed herein are compositions of retroviruses and methods of using the same for gene delivery, wherein the retroviruses comprise a viral envelope protein comprising at least one mutation that diminishes its native function, a non-viral membrane-bound protein comprising a membrane-bound domain and an extracellular targeting domain.

Helper innate lymphoid cells (ILC) play important functions in immunity and tissue homeostasis, but their development remains poorly understood. In this study, the inventors investigated the precise function of the transcription factor NFIL3, which they established as the earliest requirement during ILC development in mouse bone marrow. Using NFIL3 retroviral ectopic expression in all-lymphoid progenitors, they showed that NFIL3 triggers a developmental process that recapitulates in vitro natural ILC development, in the absence of any other instructive signal. The present invention relates to an in vitro method for obtaining innate lymphoid precursors cells, said method comprising the step of i) culturing hematopoietic precursors cells in an appropriate culture medium, and ii) manipulating the said hematopoietic precursors cells to increase the expression or function of the Nuclear Factor Interleukin (3) Regulated (NFIL3).

The present invention relates to the production, modification and use of extracellular vesicles (EVs), in particular for the delivery of payloads of nucleic acids. The EV may contain a retroviral capsid and may be coated with a polymer. The EV may be treated with a protease to remove proteins on the outer surface of the EV to form a cleaved EV. The cleaved EV may be coated with a polymer and used to deliver a payload of nucleic acid to a cell.

The present disclosure provides nanostructures and nanostructure-based vaccines. Some nanostructures of the present disclosure display antigens capable of eliciting an immune response to infectious agents such as bacteria, viruses, and pathogens. Some vaccines of the present disclosure are useful for preventing or reducing the severity of infection by infectious agents including, but not limited to, Lyme disease, pertussis, herpes virus, orthomyxovirus, paramyxovirus, pneumovirus, filovirus, flavivirus, reovirus, retrovirus, meningococcus, or malaria. The antigens can be attached to the core of the nanostructure non-covalently or covalently, including as a fusion protein or by other means disclosed herein. Multimeric antigens can optionally be displayed along the symmetry axis of the nanostructure. Also provided are proteins and nucleic acid molecules encoding such proteins, vaccine compositions, and methods of administration.

This invention provides a method for producing engineered virus-like particles using stable cell lines, and the engineered virus-like particles thereof. The method comprises the following steps: constructing a stable BaEV retrovirus packaging cell line; transfecting the BaEV retrovirus packaging cell line obtained in step 1 with a Gag-Cas9 plasmid; constructing a self-inactivating delivery plasmid for a retrovirus packaging system; and selecting a stable cell line capable of producing engineered virus-like particles. This allows for the production of engineered virus-like particles through stable cell lines. This invention innovatively develops a method for preparing eVLP stable cell lines. This method overcomes existing production bottlenecks, providing a stable, efficient, and economical gene-editing vector solution for research and clinical applications, and promoting further development in the field of gene therapy.

The present application provides a method for batch preparation of immune cell therapy products without electric conversion, the CD7 gene of the immune cells is knocked out and a chimeric antigen receptor targeting CD7 is expressed at the same time, the method comprises producing engineered viroplasm by a stable cell line, knocking out the CD7 gene in the immune cells by using the engineered viroplasm, and transducing the immune cells by using a CD7 CAR retrovirus vector, thereby obtaining the immune cell therapy product. The present application aims at the difficulties in the current CAR-T cell preparation, such as high cost, poor popularity, and high risk of failure in the application of CRISPR electric conversion technology, introduces eVLP technology for efficient and convenient gene editing, and ensures safety, saves cost, reduces operation complexity, and greatly improves the success rate of immune cell preparation. Based on clinical needs, the present application uses a CD7 eVLP vector combined with a CD7 CAR retrovirus to construct a CD7 KO+CD7 CAR T cell therapy product.

To provide retroviral and lentiviral vectors.SOLUTION: The invention provides a retroviral or lentiviral vector having a viral envelope comprising: (i) a mitogenic T cell activating transmembrane protein comprising a mitogenic domain and a transmembrane domain; and / or (ii) a cytokine-based T cell activating transmembrane protein comprising a cytokine domain and a transmembrane domain, wherein the mitogenic or cytokine-based T cell activating transmembrane protein is not part of the viral envelope glycoprotein. When cells such as natural killer cells are transduced with such viral vectors, they are simultaneously activated by the mitogenic T cell activating transmembrane protein and / or the cytokine-based T cell activating transmembrane protein.SELECTED DRAWING: None

The invention belongs to the technical field of biology, and particularly discloses a cell line capable of stably expressing an nsP4 gene and application of the cell line in preparation of a replication-deficient chikungunya virus. The inventor uses retrovirus for expressing CHIKV nsP4 to infect hamster kidney cells (hamster kidney cells), after puromycin screening, verification of passage stability of monoclonal cells, protein expression stability, replication-deficient virus titer and the like is carried out, finally, a cell line which can be used for continuous passage of CHIKV replication-deficient viruses and has high titer is obtained, and the preservation number is CCTCC NO: C2025281. The replication-deficient virus CHIKV-delta nsP4 generated by the RNA of the nsP4 gene deleted from the trans-complementary CHIKV of the cell line has good safety, immunogenicity and immune protection efficacy, can effectively resist the challenge infection of wild viruses, and can be used for preparing a safe and effective CHIKV novel vaccine candidate strain.

The present application relates to the field of cell reprogramming, and in particular to a method for inducing fibroblast transdifferentiation into kidney epithelial cells by using a double-stranded retrovirus vector combined with a double selection antibiotic screening strategy. The present application first applies a double-stranded retrovirus vector, and adds a Puromycin and Blasticin double-antibiotic screening strategy, to induce four transcription factors Hnf1beta, Emx2, Pax8 and Hnf4alpha to reprogram embryonic fibroblasts into kidney tubular epithelial cells within two weeks. The establishment of the technical system of the present application will provide strong technical support for obtaining kidney tubular epithelial cells in vitro.

Provided herein are engineered nucleic acids (e.g., expression vectors, including viral vectors, such as lentiviral vectors, adenoviral vectors, AAV vectors, herpes viral vectors, and retroviral vectors) that encode OCT4; KLF4; SOX2; or any combination thereof that are useful, for example, in inducing cellular reprogramming, tissue repair, tissue regeneration, organ regeneration, reversing aging, or any combination thereof. Also provided herein are recombinant viruses (e.g., lentiviruses, alphaviruses, vaccinia viruses, adenoviruses, herpes viruses, retroviruses, or AAVs) comprising the engineered nucleic acids (e.g., engineered nucleic acids), engineered cells, compositions comprising the engineered nucleic acids, the recombinant viruses, engineered cells, engineered proteins, chemical agents that are capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, an engineered protein selected from the group consisting of OCT4; KLF4; SOX2; or any combination thereof, an antibody capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, and methods of treating a (e.g., ocular disease), preventing a disease (e.g., ocular disease), regulating (e.g., inducing or inducing and then stopping) cellular reprogramming, regulating tissue repair, regulating tissue regeneration, or any combination thereof).

This invention discloses a nucleic acid molecule encoding a modified retroviral Gag protein and its applications. The Gag protein contains an engineered acylated tag sequence at its N-terminus, comprising: a glycine residue at the second position after the initiating methionine for myristylation modification; and / or a cysteine ​​residue in the N-terminal region for palmitoylation modification. The tag sequence is selected from SEQ ID NO:1, 3-9, or derivative amino acid sequences having at least 80% identity and retaining function. Preferably, the modification involves replacing the first 9 amino acids at the N-terminus of the wild-type HIV-1 Gag protein with the aforementioned tag to enhance membrane anchoring and improve viral packaging efficiency. This invention also provides packaging plasmids containing this molecule, host cells, packaging systems, and preparation methods. This invention can significantly improve viral yield and titer, and is particularly suitable for overcoming the efficiency bottleneck in packaging large-load lentiviruses of 7-10 kb.

The invention discloses an engineered virus-like particle (VLP) and application of the engineered virus-like particle in mRNA (messenger ribonucleic acid) encapsulation and delivery. A capsid fusion protein is constructed by fusing a retrovirus capsid protein with an RNA binding protein MCP, and meanwhile, an MS2 aptamer specifically bound with MCP is embedded into a 3'end non-coding region of a target mRNA. When the fusion protein and the target mRNA are co-expressed in mammalian cells, the mRNA can be specifically encapsulated into the self-assembled VLP. The VLP can efficiently deliver packaged mRNA to cells and achieve protein expression, has the advantages of being high in safety, accurate in targeting and good in biocompatibility, can solve the toxicity problem of an existing delivery system, is widely applied to gene therapy, tumor immunotherapy and mRNA vaccine delivery, and is huge in clinical application potential.

The application provides application of overexpressed CDCA5 in preparation of a drug for enhancing antitumor function of T cells, and relates to the technical field of cellular immunotherapy. The method for overexpressing CDCA5 is to use a CDCA5 overexpression agent, and the CDCA5 overexpression agent is a CDCA5 overexpression vector, and the CDCA5 overexpression vector is any one of a lentivirus and a retrovirus. The application overcomes the defects of the prior art, verifies that CAR-T with overexpressed CDCA5 can significantly enhance antitumor function in vivo and in vitro, and provides a new technical direction for antitumor treatment.

The invention belongs to the technical field of biological medicines, and discloses application of an MYLK3 gene inhibitor in preparation of a medicine for treating castration-resistant prostate cancer. The biological function of the MYLK3 in the prostatic cancer is confirmed for the first time, an intervention means taking the MYLK3 as a target spot is provided, and it is clear that the MYLK3 inhibitor can improve the sensitivity of the castration-resistant prostatic cancer to an androgen receptor antagonist. The invention particularly provides a gene therapeutic drug taking the human MYLK3 gene as a therapeutic target, which is suitable for treating prostatic cancer (especially castration-resistant prostatic cancer) and can also be used in combination with other targeted drugs to improve the therapeutic effect. A reasonable RNAi target sequence is designed for a target gene, a retrovirus vector plasmid containing the target sequence is successfully constructed, the retrovirus vector plasmid has a remarkable inhibition effect on the proliferation capacity of prostate cancer cells, and meanwhile, the sensitivity of the prostate cancer cells to AR antagonists is remarkably improved.

Methods of eliminating a retrovirus from a subject utilize nanoformulated anti-retroviral compounds and gene editing agents. Compositions comprise at least one anti-retroviral compounds, at least one gene-editing agent, or combinations thereof.

Compositions of retroviruses and methods of using the same for gene delivery are disclosed, wherein the retroviruses comprise a viral envelope protein comprising at least one mutation that diminishes its native function, a non-viral membrane-bound protein comprising a membrane-bound domain and an extracellular targeting domain.