183 results about "Retrovirus" patented technology

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.

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).

The present invention provides a method for packaging a BaEV retroviral vector. The method includes constructing a stable BaEV retroviral packaging cell line, transfecting a retroviral vector plasmid containing a target sequence into a virus-producing cell line, where the virus-producing cell line is a HEK293T cell line or its derivative cell line, to harvest the transiently transfected and virus-producing retroviral vector, mixing it with the stable BaEV retroviral packaging cell line, and promoting the transduction of the retroviral vector into the stable BaEV retroviral packaging cell line by horizontal centrifugation. The present invention can achieve the packaging of a BAEV-type retroviral vector carrying a CD19CAR foreign gene plasmid, construct a BAEV-type retroviral vector-producing cell line, and produce a high-titer BAEV-type retroviral vector.

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.

Provided herein are engineered nucleic acids (e.g., expression vectors, including viral vectors, such as lentiviral vectors, adenoviral vectors, AV 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).

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).

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.

The invention relates to a design method of a CAR-T cell model targeting human TROP2 and application of the CAR-T cell model in triple negative breast cancer. The method comprises the following steps: obtaining protein sequences of a light chain and a heavy chain by utilizing a sequence of a targeting drug of the human TROP2; comparing the protein sequences of the light chain and the heavy chain with the sequence of the drug protein, and removing the same sequence part to obtain a variable protein sequence; transforming the variable protein sequence into a base sequence to respectively obtain base sequences of a light chain variable region and a heavy chain variable region, and adding a signal peptide and a connecting region to synthesize two base sequences to obtain a human TROP2 antibody sequence fragment; the preparation method comprises the following steps: synthesizing plasmids carrying a human TROP2 antibody sequence fragment by taking an SFG retroviral vector as a skeleton; and enabling the human T cells to express a target gene through transfection. In the embodiment, the nucleic acid sequence of the human TROP2 antibody as shown in SEQ ID NO.1 and SFG are used as carriers, a retrovirus transfection system is used for constructing a TROP2. CAR-T cell model, the functionality of the TROP2. CAR-T cell on the triple negative breast cancer is defined, and a new method is provided for clinical treatment of the triple negative breast cancer.

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 in the central nervous system or ex vivo. 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 disease (e.g., a neurological disease), preventing a disease (e.g., neurological disease), regulating (e.g., inducing or inducing and then stopping) cellular reprogramming, regulating tissue repair, regulating tissue regeneration, or any combination thereof.

To provide novel transduction enhancers and uses thereof.SOLUTION: The present invention relates to a method for transducing a target cell, the method comprising the step of contacting a target cell with a retroviral vector and a compound capable of enhancing transduction efficiency or a combination of such compounds, wherein the target cell is pre- and / or co-stimulated by pre- and / or co-incubation with a transduction enhancing compound or a combination of transduction enhancing compounds prior to and / or during contacting the target cell with the retroviral vector.SELECTED DRAWING: None

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.

The present invention relates to the field of genetic engineering, and in particular to a viral vector and a use of a TCR subunit. Provided is a lentiviral vector or a retroviral vector. The ability to infect a target cell by the lentiviral vector or the retroviral vector is reduced. Also disclosed is a use of a TCR subunit or a functional fragment thereof in the preparation of the lentiviral vector or the retroviral vector.

The invention provides a preparation method of a Syn-CAR-T cell, the Syn-CAR-T cell and application of the Syn-CAR-T cell, and the method comprises the following steps: constructing a stably transfected SIN-gamma-RV vector, namely a PSR vector, by using a PiggyBac transposon system, preparing a Syn CAR plasmid vector, expressing a SynNotch system by using the PSR vector, combining a GAL4 induction vector and a UAS effect vector of the SynNotch system on the PSR vector, and preparing the Syn CAR plasmid vector. Enabling the SynNotch system to be delivered by one PSR vector, and constructing a plasmid of the Syn CAR retrovirus vector; and the plasmid of the retroviral vector is used for transduction of a T cell, and the Syn-CAR-T cell is prepared. Results of the invention show that the induction efficiency of the target spots BCMA, CD38 and GPC3 is extremely low, and the target spots are not suitable for the SynNotch system of the invention; the induction efficiency of the target CD19 is highest, and the target CD19 is most suitable for being applied to the SynNotch system.

The current disclosure is directed to an immunogenic composition comprising engineered extracellular vesicles with enhanced immunity, methods of making engineered extracellular vesicles with enhanced immunity, a method of slowing or stopping a disease in a subject through the use of the disclosed immunogenic composition, and a method of immunizing a subject through administering the disclosed immunogenic composition.

The present disclosure relates generally to compositions and methods comprising recombinant integration deficient retroviral vectors where the expression of cargo(s) of interest is dependent upon a PRE in the absence of an exogenous promoter.

Provided are a packaging system for packaging a lentiviral vector or a retroviral vector, and a use of the packaging system. Specifically, the provided packaging system comprises a polynucleotide encoding a CAR binding molecule, wherein the CAR binding molecule can bind to a CAR. Pseudotransduction is reduced when a host cell is transduced by the lentiviral vector or the retroviral vector packaged by the provided packaging system, and the ability to transduce a target cell expressing an antigen decreases, thereby significantly improving the targeting performance of the lentiviral vector or the retroviral vector in the transduction of immune cells such as T cells.

The invention belongs to the technical field of indication viruses, and particularly relates to a composition and application thereof in inhibition of retrovirus inactivation. The composition provided by the invention comprises 15 mM of glycine betaine and 50 mM of fructose. The fructose can increase the half-life period of the retrovirus and the cell specific productivity, and the glycine betaine can neutralize the negative effect of the high osmotic pressure environment on the cell growth. The glycine betaine and the fructose are compounded according to a certain proportion, so that the stability of the retrovirus can be improved, the inactivation of the retrovirus can be inhibited, the inactivation rate of the retrovirus can be reduced, the specific productivity of cells can be improved, the half-life period of the virus can be prolonged, the retrovirus stock solution with high stability and long half-life period can be obtained, and the time and the cost can be reduced; therefore, the safety and effectiveness of retrovirus production are improved.

The present invention relates to Split-Intron Final Self-Inactivating (SIN) retroviral vectors which comprises a viral major splice donor (mSD) comprising mutations, a mouse mammary tumor vector long terminal repeat (MMTV-LTR) or a Type B leukemogenic virus long terminal repeat (TBLV-LTR), a eukaryotic splice acceptor (eSA), and a eukaryotic splice donor (eSD). Furthermore, the invention relation to the use and methods of uses of such vectors in gene therapy.

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.

This document relates to methods and materials involved in treating cancer. For example, methods and materials for using (a) APCs (e.g., dendritic cells) designed to release a viral vector that can infect a T cell (e.g., an infectious retroviral vector or an infectious lentiviral vector) and drive expression of an antigen receptor (e.g., a CAR) within that T cell and (b) an antigenic composition containing one or more antigens that can be presented to T cells within the mammal by APCs of the administered population and / or by other APCs within the mammal to produce dual specific CAR+ memory T cells are 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.

Engineered TCR molecules specific to retroviral peptide / HLA complexes guide direct killing and / or enable robust immune responses against HIV infected cells. In a first aspect, a TCR fusion polypeptide comprises a TCR variable alpha (Vα) domain, a TCR variable beta domain (Vβ) and a TCR constant region domain (C), wherein the TCR fusion polypeptide is specific for HIV peptides.

This invention relates to retroviral gene transfer vectors, particularly lentiviral vectors, pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, comprising a promoter and a transgene; and methods of making the same. The present invention also relates to the use of said vectors in gene therapy, particularly for the treatment of respiratory tract diseases such as Cystic Fibrosis (CF).

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 invention discloses a double-antibody sandwich ELISA kit for detecting ENTV and JSRV and application of the double-antibody sandwich ELISA kit, and belongs to the technical field of biology. The double-antibody sandwich ELISA kit contains an ELISA plate coated with a monoclonal antibody for resisting goat endemic intranasal tumor virus p27 protein, a polyclonal antibody for resisting goat endemic intranasal tumor virus p27 protein and HRP (horse radish peroxidase) labeled goat anti-rabbit IgG (immunoglobulin G). Wherein the monoclonal antibody for resisting the goat endemic intranasal tumor virus p27 protein is secreted by a hybridoma cell strain ENTV-2C3, the hybridoma cell strain ENTV-2C3 is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: C 2024412. The double-antibody sandwich ELISA kit established by the invention has the characteristics of high detection sensitivity and good specificity, can be used for detecting goat endemic intranasal tumor virus and sheep lung adenoma retrovirus, and provides a technical means for prevention and control of goat endemic intranasal tumor and sheep lung adenoma.

The new alpharetrovirus-based particles are suitable for high efficiency of transiently transducing animal cells. e.g. human or murine cells, and which efficiently introduce coding and non-coding RNA contained in the alpharetrovirus-based particles into target cells The alpharetrovirus-based particles also provide for high efficiency of the activity and / or integrity of the RNA that is introduced into the animal cells, as the particles protect the incoming RNA from degradation during entry. The transferred RNA can be of non-coding nature (e.g. single guide (sg) RNA. short-hairpin (sh) RNA. micro RNA. or long non-coding (Inc) RNA. or the RNA may encode proteins or peptides. e.g. receptors. transcription factors. cellular enzymes, antigens for use in vaccination, gene / protein therapy and / or gene editing nucleases, recombinases and transposases.

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 invention relates to a method for purifying retrovirus, and particularly, the method comprises affinity chromatography, composite mode chromatography and molecular sieve chromatography. The retrovirus purified by the method is high in titer recovery rate, excellent in impurity removal effect, low in requirement on an initial to-be-purified sample and simple in amplification of a purification process, and can be used for large-scale purification of the retrovirus, and the yield and the purity of the retrovirus meet industrial requirements.

The present invention provides a method of electrotransfection-free batch preparation of an immune cell therapeutic product, the CD7 gene of the immune cell itself being knocked out and simultaneously expressing a CD7-targeted chimeric antigen receptor, comprising the production of engineered viroid particles by a stable cell line, the knockout of the CD7 gene in the immune cell using the engineered viroid particles, and the preparation of the therapeutic product of the immune cell, the preparation of the therapeutic product of the immune cell, the preparation of the therapeutic product of the immune cell, and the preparation of the therapeutic product of the immune cell. The CD7 CAR retroviral vector is used for transducing immune cells, so that the immune cell treatment product is obtained. Aiming at the difficulties of high cost, poor universality and high failure risk of CRISPR application electrotransfection technology in current CAR-T cell preparation, the eVLP technology is introduced to carry out efficient and convenient gene editing, so that the safety is guaranteed, the cost is saved, the operation complexity is reduced, and meanwhile, the success rate of immune cell preparation is greatly improved. Based on clinical requirements, a cell therapy product of CD7 KO + CD7 CAR T is jointly constructed by combining a CD7 eVLP vector with a retrovirus of CD7 CAR.