66 results about "Viral dna" patented technology

Provided herein are non-immunogenic, circular, non-integrating, non-viral DNA vectors, and methods of using such DNA vectors, which are capable of forming one or more specialized secondary structures, for example extended cruciform structures, and express one or more therapeutic genes. These DNA vectors capable of forming one or more specialized secondary structures, for example extended cruciform structures, provide for increased nucleus entry, episomal stability, and long-term transgene expression, and have unique and highly unexpectedly reduced immunogenicity. The one or more specialized secondary structures, for example extended cruciform structures, formed by the circular DNA vectors provided herein are stabilized through the interaction of two specifically oriented repeating sequences, for example AAV-derived inverted terminal repeat (ITR) sequences, separated by a small, non-repeating sequence, for example a non-repeating sequence comprising a small bacterial origin of replication (Ori).

The present disclosure relates to circular, non-viral DNA vectors, compositions including one or more of the disclosed vectors, and methods for delivering and / or expressing one or more therapeutic genes (e.g., proteins) in mammals, e.g., human patients. In some embodiments, the present disclosure is directed to circular, non-viral DNA vectors, such as circular non-viral DNA vectors including at least two inverted repeat sequences, where the at least two inverted repeat sequences are separated by a non-repeated nucleotide sequence which is not part of the at least two inverted repeat sequences. In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element.” In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element,” but include at least a portion of a bacterial origin of replication.

To provide non-viral DNA vectors for expressing FVIII therapeutic agents and uses thereof.SOLUTION: The application describes a ceDNA vector having a linear and continuous structure for the delivery and expression of a transgene. The ceDNA vector comprises an expression cassette franked by two ITR sequences, where the expression cassette encodes a transgene encoding FVIII protein. Some ceDNA vectors further comprise a cis-regulatory element including a regulatory switch. Further provided herein are methods and cell lines for reliable gene expression of FVIII protein in vitro, ex vivo and in vivo using the ceDNA vectors.SELECTED DRAWING: None

The present disclosure relates to circular, non-viral DNA vectors, compositions including one or more of the disclosed vectors, and methods for delivering and / or expressing one or more therapeutic genes (e.g., proteins) in mammals, e.g., human patients. In some embodiments, the present disclosure is directed to circular, non-viral DNA vectors, such as circular non-viral DNA vectors including at least two inverted repeat sequences, where the at least two inverted repeat sequences are separated by a non-repeated nucleotide sequence which is not part of the at least two inverted repeat sequences. In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element.” In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element,” but include at least a portion of a bacterial origin of replication.

The present invention discloses adeno associated virus mediated Lysyl Oxidase gene constructs for Keratoconus, wherein the method for construction of recombinant AAV LOX vector comprising steps of culturing human corneal fibroblast cells, generating the rAAV LOX vector by transfection, purifying the generated rAAV LOX, extracting the viral DNA, transducing the rAAV LOX vector. The present invention discloses SEQ ID NO: 3 and SEQ ID NO: 7 of codon sequence and optimized codon sequence of LOX. The present invention enhances the expression of LOX, fibronectin and Connective Tissue Growth Factor and decreases the expression of Extracellular Matrix degrading protein Matrix Metalloprotease 9. The AAV mediated LOX gene constructs aids in improved corneal stiffness, collagen expression, collagen crosslinking. The AAV mediated LOX gene constructs are safe and biocompatible.

Provided herein are lipids having the Formula I or Formula Ia:and pharmaceutically acceptable salts thereof, wherein R′, R1, R2, R3, R4, R5, R6a, R6b, X1, X2, and n are as defined herein for Formula I and Formula Ia, respectively. Also provided herein are lipid nanoparticle (LNP) compositions comprising lipid having the Formula I or Ia and a capsid-free, non-viral vector (e.g., ceDNA). In one aspect of any of the aspects or embodiments herein, these LNPs can be used to deliver a capsid-free, non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ, and the like).

The present disclosure relates to circular, non-viral DNA vectors, compositions including one or more of the disclosed vectors, and methods for delivering and / or expressing one or more therapeutic genes (e.g., proteins) in mammals, e.g., human patients. In some embodiments, the present disclosure is directed to circular, non-viral DNA vectors, such as circular non-viral DNA vectors including at least two inverted repeat sequences, where the at least two inverted repeat sequences are separated by a non-repeated nucleotide sequence which is not part of the at least two inverted repeat sequences. In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element.” In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element,” but include at least a portion of a bacterial origin of replication.

The application describes ceDNA vectors having linear and continuous structure for delivery and expression of a transgene. ceDNA vectors comprise an expression cassette flanked by two ITR sequences, where the expression cassette encodes a transgene encoding FVIII protein. Some ceDNA vectors further comprise cis-regulatory elements, including regulatory switches. Further provided herein are methods and cell lines for reliable gene expression of FVIII protein in vitro, ex vivo and in vivo using the ceDNA vectors. Provided herein are method and compositions comprising ceDNA vectors useful for the expression of FVIII protein in a cell, tissue or subject, and methods of treatment of diseases with said ceDNA vectors expressing FVIII protein. Such FVIII protein can be expressed for treating disease, e.g., hemophilia A.

Provided herein are methods and compositions for editing the genome of a cell. In some embodiments, a nucleotide sequence of at least 200 nucleotides in length is inserted into a target region in the genome of a cell.

The invention discloses a quadruple real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primer probe combination for detecting African swine fever virus and application. The primer probe combination comprises a first primer probe combination, a second primer probe combination, a third primer probe combination and a fourth first primer probe combination, the primer probe is capable of specifically targeting ASFV p72 gene, CD2v-I gene, CD2v-II gene and I177L gene, and the detection kit developed based on the primer probe combination can rapidly diagnose African swine fever virus and identify CD2v gene I and II type and CD2v and I177L gene deleted strains. The kit has extremely high sensitivity, the lowest detection limit can reach 8 copies / mu L, it is ensured that trace virus DNA can be detected, and missing detection is avoided.

The present application relates to a kit and method for the co-extraction of viral DNA and RNA, and belongs to the field of biotechnology. The present invention can directly extract viral nucleic acids (DNA and RNA) from cell-free body fluids of humans and animals, such as serum, plasma, and centrifugal supernatants of tissue grinding fluid, by combining solutions such as lysate and rinse solution with extraction methods. The method and kit of the present invention can simultaneously extract DNA and RNA, and are simple to operate, especially for small virus nucleic acid extraction with high sensitivity and better extraction effect.

The present disclosure relates to circular, non-viral DNA vectors, compositions including one or more of the disclosed vectors, and methods for delivering and / or expressing one or more therapeutic genes (e.g., proteins) in mammals, e.g., human patients. In some embodiments, the present disclosure is directed to circular, non-viral DNA vectors, such as circular non-viral DNA vectors including at least two inverted repeat sequences, where the at least two inverted repeat sequences are separated by a non-repeated nucleotide sequence which is not part of the at least two inverted repeat sequences. In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element.” In some embodiments, the disclosed circular, non-viral DNA vectors do not include a “DD element,” but include at least a portion of a bacterial origin of replication.

A solid or semi-solid state anti-viral lip balm composition comprising a direct or indirect nucleoside analog inhibitor of herpes simplex viral DNA polymerase or a pharmaceutically acceptable salt thereof mixed with at least one wax, at least one carrier oil selected from the group consisting of coconut oil including refined coconut oil and sunflower oil and a butter or butter substitute as a softening agent.

Disclosed herein are DNA amplification methods for quantifying DNA fragments of a target DNA in a sample by size. This can be used, for example, to detect tumor-derived viral DNA in blood sample and distinguish it from larger viral DNA from non-tumor sources. In particular, disclosed herein are methods of detecting, monitoring or treating a human papilloma virus (HPV)-associated malignancy in a subject that involves detecting a presence or absence of at least one circulating tumor-derived HPV DNA in a sample from the subject. Kits for accomplishing the same are also provided.

Provided herein are circular, non-integrating, non-viral DNA vectors which express a TNALP polypeptide in an improved expression cassette, wherein the DNA vectors are capable of forming one or more specialized secondary structures, for example an extended cruciform structure, and methods of using such DNA vectors. These DNA vectors having improved expression cassettes provide for increased potency and long-term TNALP transgene expression.

A recombinant lentiviral vector comprising a coding sequence for an OR protein fused to a coding sequence for a fluorescent protein or a subunit of a fluorescent protein, and a promoter active in human cells operatively linked to the coding sequences. A recombinant lentivirus comprising a recombinant genome comprising an RNA that generates an ANCH sequence upon retrotranscription. A recombinant eukaryotic cell comprising a genomically integrated DNA copy of the recombinant lentiviral vector. A method of observing lentiviral DNA in a eukaryotic cell, comprising: providing a recombinant eukaryotic cell that produces a fusion protein comprising an OR protein, fused to a fluorescent protein or a subunit of a fluorescent protein: infecting the recombinant eukaryotic cell with a recombinant lentivirus comprising a recombinant genome comprising an RNA that generates an ANCH sequence upon retrotranscription, under conditions sufficient for reverse transcription of the recombinant lentiviral genome comprising an ANCH sequence: allowing the OR protein to bind to the ANCH sequence; and detecting the fluorescent protein or subunit of the fluorescent protein to thereby observe the lentiviral DNA in the eukaryotic cell. This tool can be suitable also for in vivo applications (e.g. humanized mice) as well as for screening of new antiretroviral compounds. The HIV-1 ANCHOR system can be extended to the study of other viruses or for the screening of antiviral compounds, e.g. against SARS-COV2.

Provided herein are improved DNA vectors (e.g., circular DNA vectors and / or non-viral DNA vectors) that can provide features including enhanced expression, persistence, safety, and manufacturability. Also provided herein are methods of producing such DNA vectors, host cells (e.g., engineered bacterial cells useful for producing such DNA vectors) containing such DNA vectors, methods of using such DNA vectors (e.g., engineered bacterial cells useful for producing such DNA vectors), and methods of using such DNA vectors. Methods of expressing therapeutic sequences in target cells by administering such DNA vectors and methods of treating diseases or conditions by administering such DNA vectors), as well as pharmaceutical compositions containing such DNA vectors.

Cell-free DNA molecules in a mixture of a biological sample can be analyzed to detect viral DNA. Methylation of viral DNA molecules at one or more sites in the viral genome can be determined. Mixture methylation level(s) can be measured based on one or more amounts of the plurality of cell-free DNA molecules methylated at a set of site(s) of the particular viral genome. The mixture methylation level(s) can be determined in various ways, e.g., as a density of cell-free DNA molecules that are methylated at a site or across multiple sites or regions. The mixture methylation level(s) can be compared to reference methylation level(s), e.g., determined from at least two cohorts of other subjects. The cohorts can have different classifications (including the first condition) associated with the particular viral genome. A first classification of whether the subject has the first condition can be determined based on the comparing.

Lipids having Formula (I): (SEE FORMULA) and pharmaceutically acceptable salts thereof are provided herein, wherein R1, R2, a and b are as defined herein. Lipid nanoparticle (LNP) compositions comprising lipids having Formula (I) and a capsid-free, non-viral vector (e.g., eDNA) are also provided herein. In any aspect or embodiment hereof, these LNPs may be used to deliver a capsid-free, non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ, and the like).

The present disclosure relates to cyclic non-viral DNA vectors, compositions comprising one or more of the disclosed vectors, and methods of delivering and / or expressing one or more therapeutic genes (e.g., proteins) in mammals, e.g., human patients. In some embodiments, the present disclosure relates to cyclic non-viral DNA vectors, such as cyclic non-viral DNA vectors comprising at least two inverted repeats, where the at least two inverted repeats are separated by a non-repeating nucleotide sequence that is not part of the at least two inverted repeats. In some embodiments, the disclosed cyclic non-viral DNA vectors do not include "DD elements". In some embodiments, the disclosed cyclic non-viral DNA vectors do not include a "DD element", but include at least a portion of a bacterial origin of replication.

A recombinant lentiviral vector comprising a coding sequence for an OR protein fused to a coding sequence for a fluorescent protein or a subunit of a fluorescent protein, and a promoter active in human cells operatively linked to the coding sequences. A recombinant lentivirus comprising a recombinant genome comprising an RNA that generates an ANCH sequence upon retrotranscription. A recombinant eukaryotic cell comprising a genomically integrated DNA copy of the recombinant lentiviral vector. A method of observing lentiviral DNA in a eukaryotic cell, comprising: providing a recombinant eukaryotic cell that produces a fusion protein comprising an OR protein, fused to a fluorescent protein or a subunit of a fluorescent protein; infecting the recombinant eukaryotic cell with a recombinant lentivirus comprising a recombinant genome comprising an RNA that generates an ANCH sequence upon retro-transcription, under conditions sufficient for reverse transcription of the recombinant lentiviral genome comprising an ANCH sequence; allowing the OR protein to bind to the ANCH sequence; and detecting the fluorescent protein or subunit of the fluorescent protein to thereby observe the lentiviral DNA in the eukaryotic cell. This tool can be suitable also for in vivo applications (e.g. humanized mice) as well as for screening of new antiretroviral compounds. The HIV-1 ANCHOR system can be extended to the study of other viruses or for the screening of antiviral compounds, e.g. against SARS-CoV2.

Provided herein are circular, non-integrating, non-viral DNA vectors which express a TNALP polypeptide in an improved expression cassette, wherein the DNA vectors are capable of forming one or more specialized secondary structures, for example an extended cruciform structure, and methods of using such DNA vectors. These DNA vectors having improved expression cassettes provide for increased potency and long-term TNALP transgene expression.

The disclosure includes systems comprising non-viral DNA vectors encapsulated in lipid vesicles that are designed to achieve sustained therapeutically relevant expression levels in blood plasma of therapeutic proteins, polypeptides, or peptides. The system can express antibodies or antigen-binding fragments at sustained and therapeutically relevant levels. The system can be administered to a subject via local or systemic routes such as intravenous (IV) or intramuscular (IM), and repeat dosing produces an additive effect. For local administration, localized expression can be achieved. Applications span treatments for various conditions, including infectious diseases, cancers, and inflammatory diseases.

The invention discloses a method for screening antiviral targets by targeting virus DNA (deoxyribonucleic acid). The method comprises the following steps: respectively obtaining effective sgRNA (single guide ribonucleic acid) sequences targeting key DNA regions of varicella zoster virus, herpes simplex virus type I and Carbosi sarcoma related herpes virus; carrying out co-transfection on plasmids containing virus DNA and dCas9-sgRNA, and respectively carrying out immunoprecipitation enrichment on DNA of targeted varicella zoster virus, herpes simplex virus type I and Carbosi sarcoma related herpes virus to screen proteins for regulating virus replication; and identifying the protein by mass spectrum, and verifying that the method is also suitable for DNA screening of targeted human immunodeficiency virus to regulate and control the protein replication of the virus, so that the method for screening the protein replication of the virus by targeted virus DNA is established.

The invention discloses a linear non-viral DNA (deoxyribonucleic acid) vector closed by an ITR (internal transcriptase) terminal as well as a preparation method and application of the linear non-viral DNA vector. The linear non-viral DNA vector comprises the following elements: an asymmetric reverse terminal repeat ITR sequence and an expression cassette; the asymmetric inverted terminal repeat ITR sequence comprises a first ITR sequence and a second ITR sequence, the first ITR sequence and the second ITR sequence are respectively positioned at two ends of an expression cassette sequence so as to enable an expression cassette to be closed, and the expression cassette comprises a cis-regulatory element; the structures of the first ITR sequence and the second ITR sequence are the same or different. The linear non-viral DNA vector is simple to synthesize and low in immunogenicity, can be expressed for a long time, can be used for stable and reliable expression of target genes in vivo and in vitro, and provides a better expression vector for gene therapy.

Provided herein are lipids having the Formula (I):and pharmaceutically acceptable salts thereof, wherein R1, R2, a, and b are as defined herein. Also provided herein are lipid nanoparticle (LNP) compositions comprising lipid having the Formula (I) and a capsid-free, non-viral vector (e.g., ceDNA). In one aspect of any of the aspects or embodiments herein, these LNPs can be used to deliver a capsid-free, non-viral DNA vector to a target site of interest (e.g., cell, tissue, organ, and the like).

The invention relates to the technical field of real-time fluorescent PCR (polymerase chain reaction) detection and analysis, in particular to a method for detecting a rat parvovirus KRV strain through real-time fluorescent PCR and application of the method. The method comprises the following steps: (1) sample pretreatment: taking rat visceral organ tissues, cecum contents or excrement, blood, swabs and cell cultures as experimental animal detection samples; adding normal saline with the same volume to prepare homogenate, centrifuging, and collecting supernate for detection; blood and ascites are directly used for detection; (2) virus extraction: taking 200 [mu] L of the treated sample, carrying out virus extraction by adopting a FastPure Viral DNA / RNA Mini Kit (RC311-01) kit of the Norazan company, and taking a proper amount of final product sample virus as a subsequent template; and (3) carrying out real-time fluorescent quantitative PCR amplification by using the specific primer designed by the invention and a TaqMan probe, collecting a fluorescent signal, and carrying out analysis and judgment.

The present disclosure relates to circular non-viral DNA vectors, compositions comprising one or more of the disclosed vectors, and methods for delivering and / or expressing one or more therapeutic genes (e.g., proteins) in a mammal, e.g., a human patient. In some embodiments, the disclosure relates to circular non-viral DNA vectors, such as circular non-viral DNA vectors comprising at least two inverted repeat sequences, wherein the at least two inverted repeat sequences are separated by a non-repetitive nucleotide sequence that is not part of the at least two inverted repeat sequences. In some embodiments, the disclosed circular non-viral DNA vectors do not comprise a "DD element." In some embodiments, the disclosed circular non-viral DNA vectors do not comprise a "DD element," but comprise at least a portion of a bacterial origin of replication.

The present disclosure provides methods and compositions of modified oligonucleotide primer and probe combinations, structurally modified with locked nucleic acids, quenchers, and dyes, effective to detect tumor-derived Human Papilloma Virus (HPV) and tumor-derived Epstein-Barr virus (EBV) and, especially, to distinguish viral DNA derived from tumors from viral DNA derived from infectious viral particles.

The present disclosure relates generally to compositions and methods comprising recombinant integration deficient retroviral vectors where the expression of cargo(s) of interest is enhanced by the presence of elements that inhibit the silencing of viral DNA.