57 results about "Spliceosome complex" patented technology

The invention discloses a method for detecting a variable spliceosome in a third generation full-length transcriptome. The method comprises the following steps: merging original annular test sequences with joints removed to form a monomolecular transcript sequence, and screening a third generation full-length transcript sequence; comparing the third generation full-length transcript sequence with a reference genome sequence, and screening a third generation full-length transcript sequence having coverage and similarity with the reference genome sequence larger than preset thresholds; carrying out splicing false positive filtration and DNA contamination filtration on the screened third generation full-length transcript sequence; and carrying out gene annotation and variable spliceosome annotation on the filtered third generation full-length transcript sequence. An overlong read length of a third generation sequencing technology mentioned in the method disclosed by the invention is large enough to cover most RNA, the third generation full-length transcript sequence can be obtained by SMRT sequencing transcriptomes without being assembled, and a splicing structure of a gene can be effectively obtained by third generation transcriptome sequencing, and more perfect gene model annotation can be constructed.

The present invention provides methods and compositions for conferring tumor selective cell death on cancer cells expressing specific target precursor messenger RNA molecules (cancer cell selective target pre-mRNAs). The compositions of the invention include conditionally replicative adenoviruses that have been genetically engineered to express one or more pre-trans-splicing molecules (PTMs) designed to interact with one or more cancer cell target pre-mRNA and mediate a trans-splicing reaction resulting in the generation of novel chimeric RNA molecules (chimeric RNA) capable of encoding adenovirus specific protein(s). Adenovirus specific proteins include those proteins complementing an essential activity necessary for replication of a defective adenovirus. The methods and compositions of the invention may be used to target a lytic adenovirus infection to cancer cells thereby providing a method for selective destruction of cancer cells. In addition, the adenoviruses of the invention may be engineered to encode PTMs designed to interact with target pre-mRNAs encoded by infectious agents within a cell, thereby targeting selective destruction of cells infected with such agents.

Methods and compositions are provided for oligonucleotides that bind targets of interest. The targets include cells and microvesicles, such as those derived from various diseases. The oligonucleotides can be used for diagnostic and therapeutic purposes. The target of the oligonucleotides can be a member of a ribonucleoprotein or spliceosomal complex such heterologous nuclear ribonucleoprotein U (hnRNP U).

The invention provides a mass spectrometry identification method of nucleophosmin variable spliceosome and a gastric cancer diagnostic reagent kit. The method is combined by a Glu-C enzyme cutting method and a Q-TOF (quadrupole time-of flight) mass spectra method, and can be applied to the clinical diagnosis and treatment evaluation of the gastric cancer; and by the method disclosed by the invention, the rapid and reliable identification result can be acquired as compared with the traditional methods.

Methods and compositions for generating novel nucleic acid molecules through targeted spliceosome mediated RNA trans-splicing that result in expression of a apoAI protein, an apoAI variant, the preferred embodiment referred to herein as the apoAI Milano variant, a pre-pro-apoAI or an analogue of apoAI. The methods and compositions include pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA) capable of encoding apoAI, the apoAI Milano variant, or an analogue of apoAI. The expression of this apoAI protein results in protection against vascular disorders resulting from plaque build up, i.e., atherosclerosis, strokes and heart attacks.

Methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated trans-splicing that result in expression of an immunogenic polypeptide. The invention includes pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA) capable of encoding the immunogenic polypeptide, recombinant vector systems capable of expressing the PTMs of the invention, and cells expressing said PTMs. The target pre-mRNA are those encoding proteins that function in antigen uptake, antigen presentation and chaperoning. The methods of the invention encompass contacting the PTMs of the invention with a target pre-mRNA, under conditions in which a portion of the PTM is trans-spliced to a portion of the target pre-mRNA to form a chimeric mRNA molecule capable of encoding an immunogenic polypeptide.

The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated trans-splicing. The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA). In particular, the PTMs of the present invention are genetically engineered to interact with factor VIII (FVIII) target pre-mRNA so as to result in correction of clotting FVIII genetic defects responsible for hemophilia A. The compositions of the invention further include recombinant vector systems capable of expressing the PTMs of the invention and cells expressing said PTMs. The methods of the invention encompass contacting the PTMs of the invention with a FVIII target pre-mRNA under conditions in which a portion of the PTM is trans-spliced to a portion of the target pre-mRNA to form a RNA molecule wherein the genetic defect in the FVIII gene has been corrected. The methods and compositions of the present invention can be used in gene therapy for correction of FVIII disorders such as hemophilia A.

The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosome mediated RNA trans-splicing that result in expression of an apoA-1 variant, the preferred embodiment referred to herein as the apoA-1 Milano variant. The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA) capable of encoding the apoA-1 Milano variant. The expression of this variant protein results in protection against vascular disorders resulting from plaque build up, i.e., strokes and heart attacks. In particular, the PTMs of the present invention include those genetically engineered to interact with the apoA-1 target pre-mRNA so as to result in expression of the apoA-1 Milano variant. In addition, the PTMs of the invention include those genetically engineered to interact with the apoB or albumin or other specific target pre-mRNAs so as to result in expression of an apoB/apoA-1 and/or alb/apoA-1 wild type or Milano fusion protein thereby reducing apoB expression and simultaneously produce ApoA-1 function.

The invention discloses an Eriocheir sinensis EsSXL gene, an amplification primer group thereof and an amplification method. A full-length cDNA (complementary Deoxyribose Nucleic Acid) sequence of the EsSXL gene is cloned from an Eriocheir sinensis testis tissue by using the primer group provided by the invention, and the EsSXL gene comprises two variable spliceosomes EsSXL1 and EsSXL2, and lays an important foundation for the research of the important function of the function of the Eriocheir sinensis EsSXL gene in a sex differentiation process of the Eriocheir sinensis EsSXL gene.

The molecules and methods of the present invention provide a means for in vivo production of a trans-spliced molecule in a selected subset of cells. The pre-trans-splicing molecules of the invention are substrates for a trans-splicing reaction between the pre-trans-splicing molecules and a pre-mRNA which is uniquely expressed in the specific target cells. The in vivo trans-splicing reaction provides a novel mRNA which is functional as mRNA or encodes a protein to be expressed in the target cells. The expression product of the mRNA is a protein of therapeutic value to the cell or host organism a toxin which causes killing of the specific cells or a novel protein not normally present in such cells. The invention further provides PTMs that have been genetically engineered for the identification of exon/intron boundaries of pre-mRNA molecules using an exon tagging method. The PTMs of the invention can also be designed to result in the production of chimeric RNA encoding for peptide affinity purification tags which can be used to purify and identify proteins expressed in a specific cell type.

The molecules and methods of the present invention provide a means for in vivo production of a trans-spliced molecule in a selected subset of cells. The pre-trans-splicing molecules of the invention are substrates for a trans-splicing reaction between the pre-trans-splicing molecules and a pre-mRNA which is uniquely expressed in the specific target cells. The in vivo trans-splicing reaction provides a novel mRNA which is functional as mRNA or encodes a protein to be expressed in the target cells. The expression product of the mRNA is a protein of therapeutic value to the cell or host organism a toxin which causes killing of the specific cells or a novel protein not normally present in such cells. The invention further provides PTMs that have been genetically engineered for the identification of exon/intron boundaries of pre-mRNA molecules using an exon tagging method. The PTMs of the invention can also be designed to result in the production of chimeric RNA encoding for peptide affinity purification tags which can be used to purify and identify proteins expressed in a specific cell type.

The invention discloses a tumor marker and an application thereof. The tumor marker comprises a spliceosome 1 or an amino acid sequence SEQ ID NO.1 thereof, a spliceosome 2 or an amino acid sequence SEQ ID NO.2, and a spliceosome 3 or an amino acid sequence SEQ ID NO.3. (1) Three different spliceosomes of MXRA 7 show different change patterns in different tumors, the gene can be expressed in different tumors, and the MXRA 7 has different expression patterns in different stages of the tumors or during the growth and metastasis of the tumors, so that the occurrence, growth, metastasis and the like of different tumors can be determined according to the transcription or expression level of the MXRA7; (2) the MXRA 7 can be used as a target spot of tumor treatment, and a new tumor treatment strategy is provided by regulating and controlling the expression process of the MXRA 7; and (3) the marker disclosed by the invention is strong in specificity, and is suitable for multiple types of tumors.

Provided herein, inter alia, are anticancer polyketides. The uses of the polyketides described herein include treatment of cancer, for example, through regulation of the spliceosome and detection of spliceosome inhibition.

The invention discloses application of a PD-L1 (Programmed Death-Ligand 1) spliceosome B as a marker for instructing administration of PD-L1/PD1 immunotherapy. Expression of three different variable spliceosomes of PD-L1 can be observed in normal and mutant colorectal cancer cell systems, and eukaryotic expression vectors of the three spliceosomes are established and transfected into tumor cells without or with low PD-L1 expression. Tests further show that in co-culture of a tumor cell system with isoform b over-expression and immune cells, apoptosis of the immune cells is promoted, secretionof immune cell factors is inhibited, inhibition of immunoreactions in bodies of patients can be reminded, and clinical sample testing shows that isoform b of PD-L1 is a monitoring target of poor prognosis of tumor.

The invention relates to the technical field of medical biology and discloses a plasmid for enhancing cellular immune effect of a nucleic acid vaccine, which is designed based on monocyte chemoattractant protein-1 (MCP-1). An expressed sequence of a novel MCP-1 spliceosome molecule is inserted into a pcDNA-3.1 plasmid to form a pcDNA-3.1-MCP-1 plasmid, the plasmid can efficiently express the novel MCP-1 spliceosome molecule, and the expression ability of the co-transfected nucleic acid vaccine is not influenced. In an in vivo test, the plasmid and the nucleic acid vaccine are injected simultaneously to have the capacity of effectively inducing cellular immune response, namely antigenic specificity spleen interferon (IFN)-gamma positive cells are increased and target-killing capacity of spleen CD8 positive cells is improved. Therefore, the plasmid can be added into the conventional nucleic acid vaccine preparation for mixed injection so as to obviously accelerate the nucleic acid vaccine-induced cellular immune response, and the plasmid serves as a novel cellular immune adjuvant.

The invention relates to a cam rotor, specifically to a wear-resistant cam rotor. The wear-resistant cam rotor comprises a rotor body equipped with several convex sides. A spindle shaft is arranged atthe center of the rotor body. Engagement bodies are separately arranged on the convex sides. By separately arranging the engagement bodies on the convex sides, when abrasion of a wear resistant stickon the engagement body is large, the engagement bodies can be dismounted, and a brand-new wear resistant stick is placed into a groove, and then a spliceosome A and a spliceosome B are connected andfixed. By arranging the wear resistant stick on the top of the curved surface of the engagement body, friction between the wear resistant stick and a pump cavity can be reduced so as to reduce abrasion of the wear resistant stick. By arranging an elastic device at the bottom of the wear resistant stick, the wear resistant stick can greatly fit the pump cavity. By arranging a waterproof plate and agroove, a fluid can be prevented from entering the inside through a seam; and by arranging a deflector hole on the engagement body, a fluid can be thrown out through the deflector hole by centrifugalforce of the rotor.

The invention provides a method for expressing and purifying a simultaneous exons 3 and 5 deficient alpha synuclein alternative spliceosome protein belongs to the technical field of biology, and a method for expressing the simultaneously exons 3 and 5 deficient alpha synuclein alternative spliceosome protein in an E.coli body and purifying the protein. The method comprises the steps of obtaining of the cDNA of the simultaneous exons 3 and 5 deficient alpha synuclein, vector construction, induced expression of protein, GST-alpha-syndelta3delta5 purification and GST label removal. The method can be used for realizing the in-vitro expression of the simultaneous exons 3 and 5 deficient alpha synuclein alternative spliceosome protein; as a result, a foundation is laid for the in-vitro study of the gathering characteristic, cytotoxicity and transfer characteristic of the protein and the construction of an animal model of the simultaneous exons 3 and 5 deficient alpha synuclein alternative spliceosome protein, and a method is provided for deep studying the correlation of the protein to the pathogenic mechanism of Parkinson disease.