34 results about "Terminal Repeats" patented technology

The invention relates to the field of biological medicine, and particularly discloses a regulatory element combination for driving nucleic acid expression based on ITR-enhancer and application of the regulatory element combination. The regulatory element combination for driving nucleic acid expression based on an ITR-enhancer comprises at least one inverted terminal repeat (ITR) and at least one enhancer, the regulatory element combination is delivered through an AAV vector. The invention also provides a recombinant nucleic acid molecule for improving specific delivery of the target gene. The recombinant nucleic acid molecule comprises the regulatory element combination, an exogenous target gene and polyadenylic acid. The AAV vector provided by the invention can be used for driving the expression of a target gene without the action of a traditional promoter, and has the advantages of enhancer tissue specificity, short sequence (50-100bp), pathological microenvironment response and the like, thereby breaking through the capacity, targeting and dynamic limitation of the traditional AAV vector, improving the DNA (Deoxyribose Nucleic Acid) carrying capacity of the AAV vector, and simultaneously improving the expression of the target gene. The tissue and spatio-temporal expression specificity of a target gene is realized.

The invention belongs to the technical field of medicines, and discloses application of an adeno-associated virus vector in preparation of medicines for preventing and treating RSV infection and / or diseases caused by RSV. According to the invention, a recombinant adeno-associated virus vector is adopted for the first time to construct a medicine for preventing and treating RSV infection or diseases caused by RSV infection, and based on this, the invention provides a nucleic acid molecule which comprises: (a1) a first region which comprises a first adeno-associated virus inverted terminal repeat sequence, and (a2) a second region which comprises a second adeno-associated virus inverted terminal repeat sequence; and (a2) a second region comprising a gene encoding a respiratory syncytial virus antigen; the recombinant adeno-associated virus containing the nucleic acid molecule can generate better humoral immune effect and cellular immune effect, has a certain serum neutralization effect on strains of A and B subtypes of RSV, and particularly, when the antigen is a fusion protein of the respiratory syncytial virus or a fusion protein mutant of the respiratory syncytial virus, the recombinant adeno-associated virus has a certain serum neutralization effect on the strains of A and B subtypes of the RSV. The recombinant adenovirus vector containing the nucleic acid molecule can successfully realize the expression of the F protein before RSV fusion.

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.

Herein is reported a method for producing a recombinant adeno-associated viral particle preparation (rAAVp), the method comprising the steps of culturing a mammalian cell comprising an expression cassette directed against: a non-adeno-associated viral gene interposed between two AAV inverted terminal repeats (ITRs), and thereby producing the rAAVp; an adeno-associated virus rep gene; an adeno-associated virus cap gene; an adeno-associated virus E1A gene; an adeno-associated virus E1B gene; an adeno-associated virus E2A gene; an adeno-associated virus E4orf6; and an adeno-associated virus VA RNA gene, wherein the culturing is carried out at a pH value in the range of pH 7.4 to pH 7.6 and including a terminal value. The yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.4 to pH 7.6 and including a terminal value is higher than the yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.0 to pH 7.2 and including a terminal value, and the yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.4 to pH 7.6 and including a terminal value is lower than the yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.0 to pH 7.2. And the rAAVp produced by the culture at a pH value in the range of pH 7.4 to pH 7.6 and including an endvalue has a higher percentage of intact particles than the rAAVp produced by the culture at a pH value in the range of pH 7.0 to pH 7.2 and including an endvalue.

Provided herein, inter alia, are compositions for treating Human T-cell lymphotropic virus type 1 (HTLV-1) associated diseases. The compositions include a protein having a zinc finger domain capable of binding a sequence within an HTLV-1 long terminal repeat (LTR). Further provided are methods of treating HTLV-1 associated diseases in a subject in need thereof. The methods include administering to the subject the protein including the zinc finger domain, or a nucleic acid encoding the protein.

Provided is a promoter having high activity in an activated T-cell. The promoter comprises, from 5′-end to 3′-end, a CMV enhancer, an IFNγ promoter, and a long terminal repeat sequence from human T-cell leukemia virus that are connected in sequence. The promoter exhibits greater activity in an activated immune cell than the existing promoters and is low in activity or inactive in other non-immune cells.

The invention belongs to the technical field of in-vitro diagnosis, and particularly relates to an anti-mycoplasma pneumoniae antibody, a mycoplasma pneumoniae detection quality control product and a preparation method and application of the mycoplasma pneumoniae antibody and the mycoplasma pneumoniae detection quality control product. The invention provides an anti-mycoplasma pneumoniae antibody which comprises a light chain variable region and a heavy chain variable region, and the light chain variable region and the heavy chain variable region are specifically combined with a C-terminal immunodominance region of mycoplasma pneumoniae P1 protein. Wherein the C-terminal repeat region of the mycoplasma pneumoniae P1 protein is a core immunogenicity region, has high conservative property and strong immunogenicity, can induce a host to generate a specific IgM antibody in the early stage of infection, and is an ideal antibody target. The anti-mycoplasma pneumoniae antibody is obtained by screening through a phage display technology, has high affinity and high specificity, can effectively recognize the C-terminal immunodominance region of the mycoplasma pneumoniae P1 protein, and provides a reliable detection tool for early diagnosis of mycoplasma pneumoniae.

To provide adeno-associated variants, formulations and methods for pulmonary delivery.SOLUTION: The present disclosure provides variant AAV capsid proteins that confer tropism for lung cells, as well as recombinant adeno-associated viruses comprising the variant AAVs and pharmaceutical compositions comprising the same, and their use in the delivery of heterologous nucleic acids to lung cells for the treatment of lung disorders. (ii) nucleic acids comprising, from 5 ' to 3 ', (a) AAV2 terminal repeats, (b) a promoter, (c) a nucleotide sequence encoding a human cystic fibrosis transmembrane conductance regulator (CFTR) or a biologically active truncated CFTR lacking amino acids 708 to 759 of the human CFTR sequence, (d) a poly-adenylation sequence, and (e) AAV2 terminal repeats.SELECTED DRAWING: None

The invention relates to circular DNA molecule for rescuing recombinant adenoviruses comprising a recombinant adenoviral genome with two inverted terminal repeats (ITRs) flanking the genome ends, wherein at least one of the ITRs is associated with a target sequence adjacent to a PAM sequence, wherein the target sequence is configured for generating an RNA-guided DNA endonuclease-mediated DNA double strand break at the external end of or in close proximity outside the external end of the respective ITR, preferably within less than about 15 nucleotides. The invention also relates to a kit and a method for rescuing recombinant adenoviruses comprising or using a circular DNA molecule as described herein.

Provided are methods for treating an ocular neovascular disease in an individual by reducing Auction of central subfoveal thickness (CST) or central retinal thickness (CRT), comprising administering a unit dose of recombinant adeno-associated virus (rAAV) particles to an eye of the individual, wherein the rAAV particles comprise: a) a nucleic acid encoding a polypeptide comprising an amino acid sequence with at least about 95% identity to the amino acid sequence of SEQ ID NO: 35 and Ranked by AAV2 inverted terminal repeats (ITRs), and b) an AAV2 capsid protein comprising an amino acid sequence LGETTRP (SEQ ID NO: 14) inserted between positions 587 and 588 of the capsid protein, wherein the amino acid residue numbering corresponds to an AAV2 VP1 capsid protein.

A recombinant AAV (rAAV) suitable for use in treating adult onset neurodegeneration caused by granulin (GRN) haploinsufficiency, such as progranulin (PGRN)-related frontotemporal dementia (FTD), is provided. The rAAV comprises (a) an adeno-associated virus 1 capsid, and (b) a vector genome packaged in the AAV capsid, said vector genome comprising AAV inverted terminal repeats, a coding sequence for human progranulin, and regulatory sequences which direct expression of the progranulin. Also provided are a method for treating a human patient with PGRN-FTD and other adult onset neurodegeneration caused by granulin (GRN) haploinsufficiencies, comprising delivering to the central nervous system (CNS) a recombinant adeno-associated virus (rAAV) having an adeno-associated virus 1 (AAV1) capsid, said rAAV further comprising a vector genome packaged in the AAV capsid, said vector genome comprising AAV inverted terminal repeats, a coding sequence for human progranulin, and regulatory sequences which direct expression of the progranulin.

The present invention relates to a novel retroviral vector comprising a 5'long terminal repeat (LTR), a primer binding site, a heterologous intron, a transgene insertion site and a 3 'LTR wherein the heterologous intron is located immediately downstream of the 5' LTR. This vector has high titer productivity and is particularly suitable for gene therapy.

Herein is reported a method for producing recombinant adeno-associated viral particle preparation (rAAVp) comprising the step of cultivating a mammalian cell comprising expression cassettes for a non-adeno-associated viral gene, which is interspaced between two AAV inverted terminal repeats (ITRs), an adeno-associated virus rep gene, an adeno-associated virus cap gene, an adeno-associated virus E1A gene, an adeno-associated virus E1B gene, an adeno-associated virus E2A gene, an adeno-associated virus E4orf6 and an adeno-associated virus VA RNA gene, and thereby producing the rAAVp, wherein the cultivating is at a pH value in the range of and including pH 7.4 to pH 7.6. The yield of the rAAVp produced by the cultivating at a pH value in the range of and including pH 7.4 to pH 7.6 is higher than the yield of a rAAVp produced by a cultivating at a pH value in the range of and including pH 7.0 to pH 7.2 and the rAAVp produced by the cultivating at a pH value in the range of and including pH 7.4 to pH 7.6 has a higher percentage of full particles than a rAAVp produced by a cultivating at a pH value in the range of and including pH 7.0 to pH 7.2.

A herpes virus vector is provided with both transcriptional and translational control. Within various embodiments the herpes virus vector is based upon a modified herpes virus and has both ICP27 and ICP34.5 under control of a CEA promoter and miRNA-124 / 143, respectively, and deletion of at least one copy of terminal repeat long region is provided to increase safety without sacrificing efficacy. The herpes virus vector can also incorporate a virus-expressed cytokine cassette encoding IL-12, IL-15 / IL-15RA under the control of CXCR4 promoter.

According to various aspects of this disclosure, the present disclosure relates to a method for treating or reducing pulmonary inflammation and / or pulmonary fibrosis in a subject in need thereof comprising intramuscularly administering to the subject a recombinant adeno-associated virus (rAAV) vector. In some aspects, the rAAV vector comprises a vector genome and an AAV capsid (e.g., an AAV1 serotype). In some aspects, the vector genome comprises AAV inverted terminal repeats (ITRs) flanking an expression construct comprising a nucleotide sequence encoding a Fibroblast growth factor 21 (FGF21) or functional fragment thereof operably linked to a ubiquitous promoter.

To provide a gene therapy for limb-girdle muscular dystrophy type 2C.SOLUTION: The disclosure relates to gene therapy vectors, such as AAV vectors, comprising a polynucleotide encoding γ-sarcoglycan (SGCG), and to methods of using such gene therapy vectors to treat subjects suffering from a muscular dystrophy, e.g., limb-girdle dystrophy type 2C (LGMD2C). Provided in one aspect is a method of treating γ-sarcoglycanopathy in a subject, the method comprising administering to the subject a therapeutically effective amount of a recombinant adeno-associated virus (AAV) vector, the rAAV vector comprising a gene expression cassette comprising a polynucleotide sequence encoding γ-sarcoglycan under transcriptional control of a promoter, where the cassette is flanked by one or more AAV inverted terminal repeats.SELECTED DRAWING: None

The disclosure relates to pharmaceutical compositions and methods of using pharmaceutical compositions comprising effective dosages of an adenoviral-based biological delivery and expression system for use in the treatment or prevention of osteoarthritis in human or mammalian joints by long-term inducible gene expression of human or mammalian interleukin-1 receptor antagonist (IL-1Ra) in synovial cells, comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (IL-1Ra), left and right inverted terminal repeats (L ITR and R ITR), the adenoviral packaging signal and non-viral, non-coding stuffer nucleic acid sequences, wherein the expression of the human or mammalian interleukin-1 receptor antagonist (IL-1Ra) gene within synovial cells is regulated by an inflammation-sensitive promoter.

A recombinant AAV (rAAV) suitable for use in treating adult onset neurodegeneration caused by granulin (GRN) haploinsufficiency, such as progranulin (PGRN)-related frontotemporal dementia (FTD), is provided. The rAAV comprises (a) an adeno-associated virus 1 capsid, and (b) a vector genome packaged in the AAV capsid, said vector genome comprising AAV inverted terminal repeats, a coding sequence for human progranulin, and regulatory sequences which direct expression of the progranulin. Also provided are a method for treating a human patient with PGRN-FTD and other adult onset neurodegeneration caused by granulin (GRN) haploinsufficiencies, comprising delivering to the central nervous system (CNS) a recombinant adeno-associated virus (rAAV) having an adeno-associated virus 1 (AAV1) capsid, said rAAV further comprising a vector genome packaged in the AAV capsid, said vector genome comprising AAV inverted terminal repeats, a coding sequence for human progranulin, and regulatory sequences which direct expression of the progranulin.

The invention discloses a circular single-stranded DNA (deoxyribonucleic acid) molecule containing an inverted repeat terminal sequence and application of the circular single-stranded DNA molecule, and relates to the technical field of biology. The annular single-stranded DNA provided by the invention can comprise two gene expression cassettes, which can be the same gene or different genes. When two same genes are inserted, the problem of wrong packaging or empty packaging is avoided no matter whether the capsid packaging is forward packaging or reverse packaging. When two different genes are inserted, the two genes can be packaged in the same capsid, so that the effect of expressing two proteins by one dose of medicine is realized. Therefore, the traditional AAV three-plasmid system can be simplified, and the wrong packaging rate of the traditional AAV three-plasmid system is reduced, so that the AAV drug yield is improved; meanwhile, pollution of non-target genes can be effectively avoided, and potential viruses of an AAV three-plasmid system are removed; and two exogenous genes can be carried at the same time for gene expression, so that wide application of the AAV vector in the field of gene therapy is promoted.

The present invention relates to methods for preparing expression vectors encoding well-tolerated and highly specific tailored recombinases that are capable of recombining asymmetric target sequences within the long terminal repeats (LTRs) of proviral DNA of a variety of retroviral strains that can be inserted into the genome of a host cell, as well as the expression vectors obtained, the cells transfected with these, the recombinases expressed and pharmaceutical compositions comprising said expression vectors, cells and / or recombinases. The pharmaceutical compositions can be used, for example, for the treatment and / or prophylaxis of retroviral infections, in particular HIV infections. In particular, the present invention relates to well-tolerated and highly specific tailored recombinases that are capable of combining asymmetric target sequences in more than 90% of HIV-1 strains, thereby excising HIV-1 sequences and expression vectors encoding them.

The invention relates to the technical field of microorganisms and application thereof, and discloses a piggyBac transposon system, a recombinant vector, a gene editing method, a recombinant strain and application of the piggyBac transposon system, the piggyBac transposon system contains piggyBac transposase, an inverted terminal repeat sequence, a target load gene sequence and a selective marker gene, the vector gene is a functional marker gene and / or a reporter gene, and the expression of the transposase is controlled through a promoter PtrpC; wherein the amino acid sequence of the piggyBac transposase is as shown in SEQ ID NO. 1; and the piggyBac transposase system also contains a vector plasmid. The gene editing method of the piggyBac transposon system can be suitable for efficiently and accurately inserting exogenous genes into gibberella zeylanica, traceless and residue-free genome modification is realized, the problem of exogenous gene residue is avoided, and the method has relatively good industrial value.

USE OF ADENO-ASSOCIATED VIRUS VECTOR IN PREPARATION OF MEDICAMENT FOR PREVENTING AND TREATING RSV INFECTION AND / OR RSV-INDUCED DISEASE For the first time, a recombinant adeno-associated virus vector is used to construct a medicament for preventing and treating RSV infection or a disease caused thereby. On this basis, a nucleic acid molecule is provided, comprising: (a1) a first region comprising a first adeno-associated virus inverted terminal repeat sequence, and (a2) a second region comprising a gene encoding a respiratory syncytial virus antigen. A recombinant adeno-associated virus comprising the nucleic acid molecule can produce a relatively good humoral immune effect and a relatively good cellular immune effect, and has a certain serum-neutralizing effect on both the subtype A strain and the subtype B strain of RSV. In particular, when the antigen is a fusion protein of the respiratory syncytial virus or a mutant of the fusion protein of the respiratory syncytial virus, the recombinant adenovirus vector comprising the nucleic acid molecule can successfully achieve the expression of the RSV pre-fusion F protein.

The present disclosure provides production of a virus-derived construct plasmid. In one aspect, the present disclosure provides a plasmid encoding at least some elements of a virus-derived construct. In one embodiment, the present disclosure provides a plasmid containing at least some of nucleic acid sequences required to construct a virus-derived construct in an operably linked form. In one embodiment, the nucleic acid sequence required to construct a virus-derived construct includes (A) a nucleic acid sequence including a terminal repeat, (B) a nucleic acid sequence encoding a packaging factor, (C) a nucleic acid sequence encoding a structural protein, and (D) a nucleic acid sequence encoding a functional cofactor.

An HSV vector with an expression cassette encoding an immune cell engager protein under control of an EF1α promoter. ICP27 under control of a CEA promoter. ICP34.5 under control of miRNA-124 / 143, and ICP47 under control of a ICP27 promoter is disclosed. The HSV vector also includes deletion of one copy of terminal repeat long region and one copy of terminal repeat short region. The immune cell engager protein comprises a T-cell binding domain and a tumor-associated antigen binding domain.

In some aspects, cardiac-specific expression cassettes are provided herein. In some aspects, provided herein is an expression cassette comprising a polynucleotide sequence encoding a gene product for therapy of a heart disease, wherein the polynucleotide sequence is operably linked to promoter (e.g., a cardiac-specific promoter), and optionally an enhancer (e.g., a cardiac-specific enhancer). In some aspects, the disclosure provides recombinant adeno-associated virus (rAAV) virions, comprising a capsid protein and a viral genome comprising an expression cassette comprising a polynucleotide sequence encoding a therapeutic gene product, e.g., dwarf open reading frame (DWORF) polypeptide, operably linked to a promoter, the expression cassette flanked by inverted terminal repeats. The disclosure further provides pharmaceutical compositions and methods of treating or preventing heart disease.

The present invention relates to nucleic acid molecules and uses thereof, in particular to nucleic acid molecules comprising a first reverse terminal repeat (ITR), a second ITR, and a gene cassette encoding miRNA and / or a therapeutic protein. In certain embodiments, the therapeutic protein comprises a blood coagulation factor, e.g., a FVIII polypeptide, a FIX polypeptide, or a fragment thereof. In some embodiments, the first ITR and / or the second ITR are / is an ITR of a non-adeno-associated virus (AAV). The invention also provides a method of treating a hemorrhagic condition, such as hemophilia, comprising administering to a subject the nucleic acid molecule or the polypeptide encoded thereby.

Provided are methods for treating an ocular neovascular disease in an individual, comprising administering a unit dose of recombinant adeno-associated virus (rAAV) particles to an eye of the individual, wherein the rAAV particles comprise: a) a nucleic acid encoding a polypeptide comprising an amino acid sequence with at least about 95% identity to the amino acid sequence of SEQ ID NO: 35 and flanked by AAV2 inverted terminal repeats (ITRs), and b) an AAV2 capsid protein comprising an amino acid sequence LGETTRP (SEQ ID NO: 14) inserted between positions 587 and 588 of the capsid protein, wherein the amino acid residue numbering corresponds to an AAV2 VP1 capsid protein.

A vector composition containing i) a first vector that comprises a double-stranded cargo nucleic acid, wherein the double-stranded cargo nucleic acid comprises a first expression cassette of a gene of interest, a second expression cassette of a selectable marker, a ubiquitous chromatin opening element (UCOE) arranged upstream of and right before the first expression cassette or the second expression cassette, and a pair of transposon derived inverted terminal repeats (ITRs), wherein the first expression cassette, the second expression cassette and the UCOE are operably connected and flanked by the pair of transposon derived ITRs, and ii) a second vector that encodes and expresses a transposase, wherein the transposase is able to recognize the pair of transposon derived ITRs in the first vector and transpose the double-stranded cargo nucleic acid.

The present invention is directed to novel production methods for recombinant adeno-associated viral (AAV) vectors or AAV virus-like particles for use in research as well as therapeutic applications. In particular, the invention discloses an advantageous method for producing recombinant adeno-associated virus (rAAV) vectors or AAV virus-like particles wherein a cell is infected with a first recombinant adenoviral vector encoding the AAV Rep proteins, and a second recombinant adenoviral vector encoding the AAV Cap proteins. The cell is further provided with a transgene-vector harboring an expression cassette flanked by AAV inverted terminal repeats (ITRs) comprising a transgene. The invention further discloses a kit or a composition for producing an rAAV vector according to the method of the invention that comprises the transgene vector, the first recombinant adenoviral vector encoding the AAV Rep proteins, and the second recombinant adenoviral vector encoding the AAV Cap proteins. The method, kits and composition of the invention allow for an increased production of AAV vector particles in a stable system.

The application relates to the field of biological medicine, and particularly discloses a regulatory element combination based on ITR-enhancer-driven nucleic acid expression and application thereof. The regulatory element combination based on ITR-enhancer-driven nucleic acid expression comprises at least one inverted terminal repeat (ITR) and at least one enhancer; and the regulatory element combination is delivered through an AAV vector. The application also provides a recombinant nucleic acid molecule for improving target gene-specific delivery, which comprises the above regulatory element combination, an exogenous target gene and a poly A. The AAV vector provided by the application can drive the expression of a target gene without the action of a traditional promoter, and has the advantages of enhancer tissue specificity, a short sequence (50-100 bp) and pathological microenvironment response, thereby breaking through the limitations of traditional AAV vector capacity, targeting and dynamics, improving the DNA load of the AAV vector, and realizing the tissue and space-time expression specificity of the target gene.