41 results about "Minicircle" patented technology

The present invention relates to a plasmid for minicircle production, a method for providing a minicircle and a minicircle produced by said method as well as a pharmaceutical composition comprising the same.

The present invention relates to the production and use of covalently closed circular (ccc) recombinant DNA molecules such as plasmids, cosmids, bacterial artificial chromosomes (BACs), bacteriophages, viral vectors and hybrids thereof, and more particularly to vector modifications that improve expression of said DNA molecules.

The present invention provides a bispecific antibody, a method and applications thereof. The bispecific antibody has a first antigen binding domain capable of specifically binding to human CD3 and a second antigen binding domain capable of specifically binding to GPC3 antigen, can specifically recognize and bind to GPC3 antigen and GPC3 overexpression tumor cells including liver cancer and other GPC3-positive tumor diseases, can further bind to T cell receptors, and can mediate T cells to provide killing effects, such that GPC3 infections can be cleared, and great application values can be provided in diagnosis, treatment, prevention and detection of GPC3-related diseases. The present invention further provides minicircle DNA having a nucleotide sequence encoding the bispecific antibody. According to the present invention, the bispecific antibody and the minicircle DNA can be used for preparing antibody pharmaceutical compositions or GPC3-positive tumor cell diagnosis reagents.

Methods for the production of supercoiled DNA minicircles and their use as substrates are described herein.

The present invention relates to a method for the production of a minicircle. In the method of the present invention, a parent plasmid is provided which has a nucleic sequence flanked by recombination sites. This parent plasmid is exposed to an enzyme which causes recombination at the recombination sites, thereby to form a (i) minicircle comprising the nucleic acid sequence and (ii) a miniplasmid comprising the remainder of the parent plasmid. One recombination site is modified at the 5′ end such that its reaction with the enzyme is less efficient than the wild type site, and the other recombination site is modified at the 3′ end such that its reaction with the enzyme is less efficient than the wild type site, and the other recombination site is modified at the 3′ end such that its reaction with the enzyme is less efficient than the wild type site, both modified sites being located in the minicircle after recombination. This favours the formation of minicircle.

The invention belongs to the optical field and the field of micro-nano systems, and particularly relates to a preparation method of an optical waveguide type minicircle resonant cavity humidity sensorcoated with a TiO2 thin film. The optical waveguide type minicircle resonant cavity humidity sensor includes photoetching on an annular resonant cavity, straight waveguide and input and output grating and coating on the TiO2 thin film on a waveguiding structure. By means of the unique ambulatory mode effect of a minicircle resonant cavity prepared on the SOI waveguide, an optical wave performs resonance in a minicircle, the stable resonance phenomenon can be observed by means of a spectrometer, when the relative humidity of the external world changes, a humidity sensitive thin film absorbs water, the resonant cavity refractive index changes, and then an output spectral line observed on the spectrometer performs wave length shift, and measurement of the humidity is completed. Compared withan ordinary humidity sensor, the optical waveguide type minicircle resonant cavity humidity sensor has the advantages of being easy to operate, low in cost, small in volume, short in reaction time, high in flexibility and not prone to electromagnet interference, and therefore the sensor has the extremely high application value.

The method disclosed herein describes a novel technology offering unparalleled efficiency, flexibility, utility and speed for the stable integration of transgenes into lymphocytes and other mammalian cells. The novel method is based on the use of an mRNA-encoded transposase (e.g. sleeping beauty transposase) in combination with a minicircle DNA-encoded transposable element. The novel method enables higher gene-transfer rates and is at the same time less toxic than the conventional approach, which is the use of plasmid DNA-encoded transposase in combination with a plasmid DNA-encoded transposable element. Applications of the invention include but are not limited to the stable integration of a transgene encoding an immune receptor (e.g. a T-cell receptor or synthetic chimeric antigen receptor) into human T lymphocytes, with the immune receptor conferring specificity for a molecule expressed by a tumor cell. The transposase mRNA and transposon minicircle DNA may be introduced into lymphocytes by methods including but not limited to electrotransfer such as electroporation and nucleofection.

The invention relates to a minicircle transfer vector for producing viral vectors comprising a transfer sequence and specific packing signals flanking both sides of the transfer sequence for packaging of the transfer sequence into particles of a viral vector. The invention also relates to minicircle packaging vectors carrying support functions for producing viral vectors. The invention further relates to cells bearing the disclosed minicircles. The invention further relates to methods for producing viral vectors using such minicircles and viral vectors obtained thereby, as well as kits useful in performing the described methods.

The invention discloses a minicircle gene vector and its preparation method and use. The minicircle gene vector does not contain a backbone DNA ingredient which can close transgene expression and is in a standard plasmid. The minicircle gene vector comprises a promoter, an ingredient for gene expression and virus replication in a viral genome, and a transgene or a transgene expression box for expression of a treatment transgenosis product. The ingredient for gene expression and virus replication in a viral genome is cloned into a vector so that the combined chimeric minicircle gene vector is obtained. After being fed into a target cell infected by viruses, a small amount of the minicircle gene vectors can produce recombinant viruses having treatment and infection effects. Through infecting a new target cell, the recombinant viruses produce new recombinant viruses having infection effects, and finally, the recombinant viruses diffuse to all target cells. Compared with the traditional gene vectors, the minicircle gene vector can be efficiently delivered into a target cell and has strong and lasting treatment effects. The invention also provides a preparation method and a use of the minicircle gene vector.

Bispecific antibodies (bsAbs) have emerged as a class of promising anti-cancer and anti-infection biological drugs. They are capable of killing target cells, either cancer cells or microbe-infected cells, at levels of nanograms per milliliter serum in vivo, about 1e+5 folds more powerful than regular antibodies. To bypass the problems of high cost in production and inconvenience in administration, a logical solution is to use gene therapy vectors to produce them in vivo. In a series of preclinical studies, we have demonstrated that DNA MiniCircle was able to express far above therapeutic levels of bsAB persistently both in the presence as well as the absence of transfection co-factors. As a specific and intended improvement of the claimed invention, an enhanced form of bispecific antibodies incorporating a target cell-effector cell bridging device (BTEC) is additionally disclosed.

Methods for the production of supercoiled DNA minicircles and their use as substrates are described herein.

The present invention provides minicircle nucleic acid vector formulations for use in administering to a subject, wherein the minicircle nucleic acid vectors include a polynucleotide of interest, a product hybrid sequence of a unidirectional site-specific recombinase, and are devoid of plasmid backbone bacterial DNA sequences. Also provided are methods of producing the subject formulations as well as methods for administering the minicircle nucleic acid vector formulations to a subject. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications.

The invention provides methods and compositions for producing and using minicircle DNA molecules that are useful for plant transformation. The invention also provides methods for transforming plant cells and plants with such minicircle DNA molecules, plant cells and plants produced by such methods, and plants transformed with minicircle DNA molecules. The methods and compositions of the invention are particularly useful for producing “intragenic plants” which do not contain any non-native DNA.

The invention belongs to the field of biomedicines and particularly relates to an immunoglobulin of yolk (IgY) of HPV (Human Papilloma Virus) L1 recombinant minicircle DNA (DeoxyriboNucleic Acid) andapplication of the IgY in preventing / treating HPV infection, wherein the HPV L1 recombinant minicircle DNA contains an L1 gene fragment of HPV16 and a signal peptide sequence, and is used as a nucleicacid antigen for directly immunizing laying hens to obtain the IgY disclosed by the invention. The biological activity of the IgY obtained by immunizing the laying hens through DNA nucleic acid antigen intramuscular injection is far superior to that of IgY prepared by a general technique (subcutaneous injection of a protein antigen), the antibody titer is higher and the specificity is higher. TheIgY disclosed by the invention can be used for preparing a drug for preventing / treating the HPV infection; a high-activity anti-HPV antibody is inputted into HPV-positive patients by vaginal medication; by combining with a nano-liposome method, an antibody transmembranely enters the cell and directly neutralizes an HPV virus in cells. The IgY disclosed by the invention is of great significance for preventing cervical cancer.

The invention belongs to the technical field of bioengineering, and particularly relates to construction and application of a model for researching interaction between HBV cccDNA and a host. The invention provides the construction and the application of the minicircle DNA model highly simulating hepatitis B virus genome cccDNA. By constructing a pmini-MC-HBV-Intron plasmid, a high-purity MC-HBV minicircle plasmid is obtained in vitro; and by optimizing a chimeric intron sequence and a secondary structure of RNA, efficient and variable splicing of the chimeric intron on the RNA level is achieved, and the MC-HBV capable of being copied at a high level in vitro is obtained. The MC-HBV can highly simulate the physiological activity between an HBV virus and host cells. The invention further discloses an in-vitro marker MC-HBV model and a detection technology, and provides the excellent model for revealing the interaction relationship between the HBV cccDNA and the host, and a function regulation mechanism, and exploring a new HBV treatment target.

The invention relates to muscle-targeted minicircle DNA gene therapy, in particular to a minicircle DNA vector for long-term efficient expression of a therapeutic gene product in muscle. The minicircle DNA vector is combined with a muscle targeted delivery technology, and can efficiently express the therapeutic gene product in muscle for a long time, thereby overcoming the defects of limited half-life period of protein drugs and unsustainable efficacy. The minicircle DNA vector can be used for treating common hereditary genetic defects or chronic diseases, and has the advantages of safety, effectiveness and low cost.

The present invention provides a bispecific antibody, a method and applications thereof. The bispecific antibody has a first antigen binding domain capable of specifically binding to human CD3 and a second antigen binding domain capable of specifically binding to CD44v6 antigen, can specifically recognize and bind to CD44v6 antigen and CD44v6 overexpression tumor cells including liver cancer and other CD44v6-positive tumor diseases, can further bind to T cell receptors, and can mediate T cells to provide killing effects, such that CD44v6 infections can be cleared, and great application valuescan be provided in diagnosis, treatment, prevention and detection of CD44v6-related diseases. The present invention further provides minicircle DNA having a nucleotide sequence encoding the bispecificantibody. According to the present invention, the bispecific antibody and the minicircle DNA can be used for preparing antibody pharmaceutical compositions or CD44v6-positive tumor cell diagnosis reagents.

The invention belongs to the field of biological medicine and particularly relates to an HPV (human papilloma virus) L1 minicircle DNA vaccine and an application of the vaccine to prevention of HPV infection. The HPV L1 minicircle DNA contains a nucleotide sequence of HPV 16 subtype L1 (major capsid protein). Meanwhile, the minicircle DNA has following advantages: 1), the minicircle DNA is not integrated into a host cell genome, cannot induce new mutation and is safer than a virus vector; 2), the minicircle DNA can be amplified massively in engineering bacteria, the preparation technology is simple, and industrial production cost is low; 3), exogenous gene expression is more stable and more efficient as compared with expression of traditional plasmid. The HPV L1 minicircle DNA has great significance for cervical cancer prevention when applied to vaccine preparation.

The application relates to a minicircle DNA expression bridging molecules for connecting human and animal target cells with effector cells, and an application of the minicircle DNA expression bridgingmolecules, in particular to a minicircle DNA vector for in vivo expression human-animal cross reaction bispecific antibody(crBsAb). The human-animal bispecific antibody(crBsAb) can combine human or animal target cells, and can also combine human or animal effector cells (T cells or natural killer cells) so as to treat cancer, infection and other diseases. Before clinical testing with medicines, the human-animal cross reaction bispecific antibody is favorable for evaluation of the safety and the validity of bispecific antibody drugs with an animal model, so as to promote development process ofthe medicines. The bridging molecules are completely the same in large animal experimentation and clinical minicircle DNA-gene drugs for human, and the condition that in the prior art, two differenteditions of biology drugs for common-used animal and human can cause difficult result interpretation, is avoided.

The present invention relates to nucleic acid molecule compositions comprising minivectors encoding a nucleic acid sequence and methods of gene therapy and prophylaxis against infection using minivectors encoding a nucleic acid sequence.

The invention discloses a detection probe, a preparation method thereof, and application, and belongs to the field of biology. According to the preparation method disclosed, full-length probe fragments are obtained through amplification by combining minicircle plasmids with a rolling circle amplification method, and the probes with different length requirements are further prepared through ultrasonic interruption. The method can be used for producing DNA probes or RNA probes, and the preparation method is simple, the yield is high, the cost is low, and the preparation period can be shortened to one day. Besides, the lengths of the probes prepared by the method is not limited, the probes with different targets and different lengths can be provided as required, the homogeneity is good, and the purity is high, so that the capture performance of the probes is greatly improved, the sequencing uniformity and depth of a target area are better, and the experiment cost is further reduced.

The invention provides application of a human hepatocyte growth factor gene in eczema treatment and a microneedle medical instrument. The human hepatocyte growth factor gene is constructed into an eukaryotic expression vector, a virus vector and a gene expression cassette or minicircle DNA (Deoxyribose Nucleic Acid), and can obviously play a role in resisting eczema and related symptoms after being locally introduced subcutaneously by adopting a microneedle or injection mode and the like. Therefore, the human hepatocyte growth factor gene has an extremely good application prospect in the aspect of eczema treatment, and especially in the form of a microneedle instrument, not only can the eczema symptom be inhibited, but also the side effect is small.

The invention relates to minicircle DNA vaccine design and application. Specifically, the invention relates to a minicircle DNA vector for expressing specific antigens or antigen fragments of pathogenic microorganisms in vivo. The minicircle DNA vector can mediate antigen proteins to be efficiently expressed in vivo, the immunogenicity of DNA vaccines is enhanced, and meanwhile, the safety problemcaused by resistance genes can be avoided. The minicircle DNA vector can be used for preventing and / or treating common infectious diseases and related cancers, and has the advantages of stronger antigen expression and higher safety.

The invention relates to MC (Minicircle) DNA (Deoxyribonucleic Acid) for expressing Her2-BTEC [Bridge between Her2 (Human Epidermal Growth Factor Receptor-2)-Positive Cells and Effector Cells] and application thereof, in particular to an MC DNA carrier for expressing an anti-Her2 bispecific antibody in vivo. Her2 positive cancer cells (target cells) are killed by bispecific antibody mediated effector cells (T cells or NK cells). The invention firstly discloses an MC DNA carrier design scheme of MC.Her2-BTEC, and the MC DNA carrier design scheme is suitable for treating cancer related to Her2.

The present invention relates to the production and use of covalently closed circular (ccc) recombinant DNA molecules such as plasmids, cosmids, bacterial artificial chromosomes (BACs), bacteriophages, viral vectors and hybrids thereof, and more particularly to vector modifications that improve expression of said DNA molecules.

The invention relates to a minicircle DNA carrier expressing IgG antibody, a preparation method and application thereof, in particular to a minicircle DNA carrier for in vivo expression of IgG antibody with complete functions. The IgG antibody neutralizes human immunodeficiency virus (HIV), other pathogene antigen or cancer cell antigen. The minicircle DNA carrier and the IgG antibody can be usedfor preventing and treating diseases, such as acquired immune deficiency syndrome (or HIV infection), other pathogen infection or cancers.

A method for the in vitro production of DNA minicircles includes steps of: a) providing nicked double-stranded oligodeoxynucleotides bunt-ended substrates having at least one phosphorylated 5′ end, b) performing a ligase-mediated circularization on a reaction mixture including the nicked double-stranded oligodeoxynucleotides substrates and a DNA bending protein, and c) obtaining DNA minicircles.

The present invention relates to a non-viral minicircle vector expressing a SOX gene, a stem cell into which the vector is introduced, a pharmaceutical composition for preventing or treating a cartilage disease, including the stem cell, and a method for constructing the vector. The transformation of mesenchymal stem cells with MC / SOX-Trio or MC / SOX-Duo, which is a non-viral minicircle vector according to the present invention, can completely exclude the necessity of expensive growth factors that have been indispensably used in inducing the differentiation of mesenchymal stem cells into chondrocytes. Accordingly, the mesenchymal stem cells transformed therewith, when implanted in vivo, can differentiate into chondrocytes by themselves, and thus have an advantage capable of simplifying the existing complicated steps of culturing cells to induce differentiation and then transplanting the cells.Further, unlike existing vector systems in which antibiotic-resistant genes and other bacteria-derived exogenous genes are simultaneously transferred to cells even after transformation, the vector of the present invention minimizes transfer of unnecessary genes into target cells by allowing two or three SOX genes necessary only for differentiation into chondrocytes to be regulated under one promoter, and thus can be utilized as a non-viral vector system in the most advantageous form for use in clinical application of stem cell-gene therapeutic agents.

Compositions and methods are provided for the treatment of an ischemic cardiovascular condition by providing a patient with a novel non-viral minicircle DNA vector comprising polynucleotide sequences that potentiate HIF-1 activity, including RNAi or antisense agents selective for proteins involved in HIF1 inactivation.

The present invention provides minicircle nucleic acid vector formulations for use in administering to a subject, wherein the minicircle nucleic acid vectors include a polynucleotide of interest, a product hybrid sequence of a unidirectional site-specific recombinase, and are devoid of plasmid backbone bacterial DNA sequences. Also provided are methods of producing the subject formulations as well as methods for administering the minicircle nucleic acid vector formulations to a subject. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications.