50 results about "Gene knockdown" patented technology

The application discloses a kind of carrier, kit and application for long-acting gene knockdown of animal parasitic nematode, belong to the field of animal parasitic disease prevention and control.The lentivirus vector in the application integrates specific coding sequence to the genome of the infected nematode, stably and continuously expresses short hairpin RNA (shRNA) in the nematode, efficiently produces primary single-stranded small RNA (siRNA), these siRNAs target homologous target gene mRNA to cause its degradation, realize stable and continuous gene silencing effect.The application breaks through the bottleneck of traditional animal parasitic nematode RNA interference (RNAi) technology with low efficiency and instability, significantly improves the timeliness of conventional RNAi, can be used to establish long-acting RNAi technology system of animal parasitic nematode, has application value in the research of host in vivo process such as nematode infection, parasitism and pathogenicity and animal parasitic nematode disease prevention and control.

The present invention relates to systems and methods for inducing and analyzing gene knockdown and / or knockout in plants at a large-scale. In particular, the present invention discloses Cell type-specific Multi-targeted AmiRNA or CRISPR (cMAC) function-scale toolbox, presenting a novel system for knockdown and / or knockout of gene families at the functional-scale, for example, at the transportome scale, and at the cell type level.

The present invention discloses a grouper GU6-1 promoter that mediates gene knockdown, and the nucleotide sequence of the grouper GU6-1 promoter is shown in SEQ ID NO: 1. Also disclosed is the use of the grouper GU6-1 promoter in mediating gene knockdown. The present invention provides a grouper U6 promoter that can efficiently mediate gene knockdown and its use in RNAi. The GU6-1 promoter can efficiently, stably, continuously and safely express shRNA to induce gene silencing, providing a reliable new tool and technical platform for problems such as gene function research, variety improvement and disease control in fish and even other animals.

The invention discloses a gene OsNST2 for transporting nucleotide sugar and application of the gene OsNST2. A nucleotide sequence of a coding region of the gene is as shown in SEQ ID No. 1, and an amino acid sequence coded by the gene is as shown in SEQ ID No. 2. It is found for the first time that the growth, development and yield of rice are affected by the OsNST2 by regulating and controlling formation of rice cell walls, an OsNST2 gene knock-down strain is created through an RNAi technology, an OsNST2 gene overexpression strain is obtained through an overexpression transgenic technology, it is verified that the OsNST2 gene is a new gene for regulating and controlling formation of the rice cell walls, and a new gene resource is provided for high yield of rice.

The invention relates to the technical field of metabolic engineering bacteria. The invention provides an engineering bacterium for improving the yield of poly-beta-hydroxybutyric acid and a construction method of the engineering bacterium. According to the invention, methylotrophic bacteria MB200 is taken as an original strain, a mispairing repair system MutL gene knock-down strain of CRISPRi is constructed, so that the mispairing repair system MutL gene knock-down strain has a higher gene mutation rate, laboratory directed evolution is carried out by utilizing a screening mode of'rich-deficient carbon sources', chassis cells with stable genomes after directed evolution are obtained, and the engineering strain is named as MB-P1. Compared with the original strain methylotrophic bacteria MB200, the PHB production capacity of the MB-P1 is improved by 72.61%. The method provided by the invention is suitable for optimizing the methylotrophic bacteria strain for producing the PHB, provides a new normal form for realizing efficient accumulation of the PHB, and has wide industrial application potential.

The invention belongs to the technical field of genetic engineering of animal molecular breeding, and particularly relates to application of egg white kinase JNK1 in resisting mycoplasma gallisepticum (MG) infection. The invention finds that the expression quantity of the JNK1 gene in chicken lung tissues and cells after MG infection is obviously reduced. Silent JNK1 can significantly inhibit replication and adhesion of MG in host cells, promote cell proliferation, inhibit release of pro-inflammatory factors and promote release of anti-inflammatory factors, which indicates that the JNK1 gene is a susceptible gene of MG, and the gene knockdown / knockout can significantly improve resistance of chicken to MG. The JNK1 gene provided by the invention can be used as a target spot for resisting replication of mycoplasma gallisepticum, can be used as a disease-resistant molecular marker, is used for carrying out auxiliary breeding on new varieties for resisting mycoplasma gallisepticum infection, and has important application value in genetic auxiliary disease-resistant breeding.

The present application provides a polypeptide targeting formyl peptide receptor on the cell membrane surface, a pharmaceutical composition and their applications. The polypeptide targeting formyl peptide receptor on the cell membrane surface includes a ligand of the formyl peptide receptor and Cys or Lys located at the C-terminus of the polypeptide connected to the ligand of the formyl peptide receptor; or, the polypeptide is the ligand of the formyl peptide receptor, and the C-terminus of the ligand of the formyl peptide receptor is Cys or Lys. After the polypeptide stimulates the formyl peptide receptor on the cell membrane surface, the endocytosed formyl peptide receptor cannot be recycled to the cell membrane, thereby targeting and rapidly knocking down the formyl peptide receptor on the cell membrane surface. Moreover, by using this polypeptide to knock down the formyl peptide receptor on the cell membrane surface, there is no need to perform enzymatic modification on the polypeptide additionally, no need to rely on other degradation techniques, and the off-target effects of gene knockout and gene knockdown methods can be avoided. The operation is simple and the effectiveness is high.

This invention belongs to the field of biomedical technology, specifically involving Trmt61a The application of genes in the preparation of drugs for treating pathological myocardial hypertrophy. This invention provides... Trmt61a The application of genes in the preparation of drugs for treating pathological myocardial hypertrophy, the aforementioned Trmt61a The nucleotide sequence of the gene is shown in SEQ ID NO.1. Animal experiments have shown that knocking down... Trmt61a Gene knockdown can significantly improve cardiac contractile function after aortic arch coarctation surgery, alleviate cardiomyocyte hypertrophy, reduce the degree of cardiac fibrosis, and effectively treat pathological myocardial hypertrophy. This invention is the first to demonstrate knockdown... Trmt61a Genes can effectively improve pathological myocardial hypertrophy, providing a new drug development path and target for the prevention and treatment of this disease, and have important medicinal value and clinical application prospects.

The present invention relates to the field of biotechnology, and discloses a novel Cas13a / cr:mi:crRNA gene knockdown tool, a construction method and an application thereof. Two regularly clustered short palindromic repeat sequences RNAa (CRISPR-Derived RNA, crRNAa) and regularly clustered short palindromic repeat sequences RNAa (CRISPR-Derived RNA, crRNAb) sequences that guide Cas13a nuclease (Cas13a) to a target gene are respectively connected to the left and right wings of a primary miRNA (pri-miRNA) to form a cr:mi:crRNA guide RNA that triple-recognizes a target gene, and constructs Cas13a / cr:mi:crRNA. After bovine and mouse muscle cells are transfected with Cas13a / cr:mi:crRNA, cr:mi:crRNA is cut and separated with the help of Drosha nuclease to produce crRNAa, crRNAb and precursor miRNA (Previous miRNA, pre-miRNA) separated from each other, thereby performing triple recognition and cutting of the targeted messenger RNA (Messenger RNA, mRNA), significantly improving the efficiency of RNAi and providing an efficient gene interference system for biological breeding.

The invention belongs to the technical field of biological medicines, and discloses an oligonucleotide molecule for targeted inhibition of a human LINC00921 gene, a coding oligopeptide and application of the oligonucleotide molecule in treatment of lung adenocarcinoma. The invention also discloses application of related drugs or products taking the oligonucleotide molecule for targeted inhibition of the human LINC00921 gene as an active ingredient in inhibition or treatment of lung adenocarcinoma. A specific siRNA target sequence is designed aiming at a target gene, a retrovirus vector plasmid containing the target sequence is successfully constructed through genetic engineering, a recombinant retrovirus capable of efficiently silencing the LINC00921 gene is obtained, the mRNA expression level of the LINC00921 gene in lung adenocarcinoma cells can be remarkably reduced, the gene knock-down effect is outstanding, and the retrovirus can be applied to lung adenocarcinoma cells. The compound has a remarkable inhibition effect on proliferation, migration and invasion ability of lung adenocarcinoma cells. Therefore, the invention has important application value in treatment of lung adenocarcinoma, and has high-throughput operability and good repeatability.

Disclosed are methods and pharmaceutical compositions for producing adjustable protein target-specific polypeptides, use for adjustable regulation of the level of one or more protein targets or a family of related protein targets and associated biological functions thereof, use for treating or preventing a disease and / or condition, selective detection and / or isolation of one or more protein targets, and use of studying the function of one or more protein targets using polypeptides that are analogs of gene knockdown by RNA interference or gene knockout by CRISPR / Cas9 in a subject.

The invention belongs to the technical field of biological medicine and molecular biology, and particularly relates to application of LGR5 in diagnosis and treatment of vascular calcification. Experiments prove that the key effect of the LGR5 in aortic smooth muscle calcification is found for the first time, it is indicated that the LGR5 is a new target spot for regulating and controlling vascular calcification, meanwhile, vascular calcification can be accurately inhibited through knockdown / knockout of the specific Lgr5 gene of vascular smooth muscle, and meanwhile, vascular calcification can be accurately inhibited through knockout / knockout of the specific Lgr5 gene of the vascular smooth muscle. According to the application disclosed by the invention, the mechanism that the Lgr5 regulates and controls the osteogenic differentiation of the vascular smooth muscle cells through the Wnt / beta-catenin signal channel is preliminarily clarified, and a clear action target and a theoretical basis are provided for drug research and development; the invention provides a new strategy for treatment of diseases such as atherosclerosis and the like, and has a wide clinical application prospect.

The present invention provides a method for constructing an animal model of systemic lupus erythematosus, belonging to the technical field of methods for constructing disease animal models. In the present invention, mice GPX4<supgt;fl / wt< / supgt;VilCre+ containing heterozygous GPX4 gene and Cre gene are first obtained, and then SLE induction drugs are used to further induce the mice to develop the disease. In view of the specific gene knockdown in specific cells in the present invention, the etiology of the disease development in the model is clear, that is, the abnormal intestinal barrier system triggers adaptive immune changes, recognizes autoantigens, and ultimately leads to the development of lupus. At the same time, the finally obtained SLE model in the present invention can exhibit higher autoantibodies, lower complement, higher inflammatory factors, and more severe lung damage, and the decrease of intestinal GPX4 in these mice is similar to the intestinal expression characteristics of human SLE.

The present application relates to the technical field of biological medicine, and in particular to a synthesis method and application of circular shRNA with gene knockdown function. The present application provides a synthesis method of circular shRNA with gene knockdown function. The ends of linear shRNA are connected by 2', 5'-phosphodiester bond to form circular shRNA, which can improve the stability of delivery in the environment of exonuclease. Moreover, after entering the cell, the 2', 5'-phosphodiester bond can be cleaved by DBR1 (Debranching RNA Lariats 1), so as to obtain shRNA, which can play the gene knockdown function of shRNA and play the role of inhibiting gene expression. The present application provides a new direction for developing a new nucleic acid drug for knocking down gene expression, and has a broad application prospect.

The invention belongs to the technical field of drug delivery systems, and particularly relates to a nucleic acid and chemotherapeutic drug co-loaded LNP delivery system and application thereof. According to the LNP delivery system, lipid nanoparticles serve as a carrier, the carrier encapsulates drug components, the encapsulation efficiency of RNA molecules and chemotherapeutic drugs in the same LNP delivery system can be effectively balanced, the administration concentration of the RNA molecules and the chemotherapeutic drugs can be met, cost is saved, introduction of new immunogens is effectively avoided, and immune risks are reduced. In addition, the invention constructs an active targeting type PEGylated lipid derivative material capable of simultaneously meeting active targeting, blood long circularity and low immunogenicity. The active targeting type LNP delivery system which is obtained by taking the PEG-lipid as the PEG-lipid of the LNP delivery system and co-carries nucleic acid and chemotherapeutic drugs remarkably improves the gene knock-down efficiency of RNA molecules and the anti-tumor activity of the chemotherapeutic drugs, can effectively overcome the problem of drug resistance of cancer chemotherapy, and is particularly suitable for cancer combined treatment.

The invention discloses a method for screening diabetes treatment targets by using drosophila melanogaster as a model organism, and belongs to the technical field of biological medicines. According to the invention, a GAL4 / UAS system is utilized to construct transgenic drosophila melanogaster of which the specificity of a target gene (X) is knocked down, and F1-generation male ppl-GAL4 / + is obtained through hybridization; uAS-X RNAi / + drosophila melanogaster; compared with a high-glucose model control group with the same background, if the X gene knocks down, lipid droplet accumulation is reduced, and circulating glucose and trehalose are reduced (both have statistical significance, Plt, 0.05), the X gene is identified as a potential diabetes (abnormal glucose and lipid metabolism) treatment target. The method provided by the invention overcomes the problems of high cost, low efficiency, insufficient individuation and the like in the existing diabetes drug screening method, and provides an accurate, rapid, efficient and low-cost screening means for research and development of diabetes treatment drugs.

The invention provides an ictalurus punctatus sex regulation target, a zbtb38 gene knockdown ictalurus punctatus sex reversal method and application, and belongs to the technical field of sex regulation. According to the invention, the sex control effective regulation target spot zbtb38 gene is successfully screened out, and the shRNA target spot is designed aiming at the coding region of the channel catfish zbtb38 gene, so that compared with CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats), the shRNA target spot has the characteristics of easiness in death and low success rate, and the expression inhibition of the channel catfish zbtb38 gene is effectively realized through site-directed gene knock-down. According to the method, the ietalurus punetaus sex reversal model is successfully established, all-female reversed fishes and hermaphrodite reversed fishes are obtained, the method has important value for sex determination mechanism research of the ietalurus punetaus, optimization of an all-male breeding strategy can be reversely guided, a scientific basis is provided for all-male group breeding and breeding cost optimization, and the method has wide application prospects. And finally, the high-quality development of the ictalurus punctatus breeding industry is promoted.

The invention discloses a BCG3973 gene knocked-down recombinant bacillus calmette guerin vaccine strain as well as a construction method and application thereof. According to the invention, a CRISPRi / dCas9th1 technology is utilized to block RNA polymerase movement in space through a compound formed by crRNA, dCas9 protein and sgRNA at a transcriptional level, so that the expression of the BCG3973 gene in the BCG vaccine is inhibited. After being treated by the recombinant bacillus calmette guerin bacterial strain, the immune-related pathways of macrophages, such as a cytokine-cytokine receptor action pathway, a Ca < + + > signal pathway and pathways such as PPAR and JAK-STAT, are obviously increased, and IL-1beta, TNF-alpha, antiviral protein and calmodulin with higher levels are expressed, so that the recombinant bacillus calmette guerin bacterial strain has stronger bactericidal ability on escherichia coli. Mouse in-vivo experiments show that the recombinant bacillus calmette-guerin bacterial strain has a very good protection effect on escherichia coli infection and endotoxemia caused by lipopolysaccharide, and shows higher survival rate and stronger capability of eliminating heterologous substance infection under attack of a lethal dose of Candida albicans.

The invention belongs to the technical field of biological medicine, and discloses application of RNF5 in preparation of medicine for preventing and / or treating acute lung injury. The invention finds that RNF5 gene knock-down significantly improves lung injury, pulmonary edema, lung inflammation and apoptosis under LPS stimulation; the RNF5 serving as a targeting gene can be used for targeted development of drugs for preventing, relieving and / or treating the acute lung injury, and has important significance on clinical treatment of the acute lung injury.

The application discloses application of an RFWD3 gene as an esophageal cancer treatment target point. The application first constructs an RFWD3 gene knockdown virus, detects the proliferation and apoptosis of esophageal cancer cells after the RFWD3 gene is knocked down at a cell level, and the result shows that knocking down the RFWD3 has the effect of inhibiting the proliferation of esophageal cancer cells and promoting the apoptosis of esophageal cancer cells. Further, the application verifies that knocking down the RFWD3 can effectively inhibit the proliferation of tumor cells and the growth process of tumors by constructing an esophageal cancer animal model in vivo. Therefore, the application provides a new target point for treating esophageal cancer, and is expected to screen and prepare a new drug for effectively treating esophageal cancer aiming at the target point.

The methods of the present disclosure include a dose-response model (DoReSeq) and machine learned models for quantifying oligonucleotide mediated off-target gene or on-target gene knockdown, and / or characterizing the level of gene expression dependent upon concentration of an oligonucleotide.

The invention provides a protein editing system and application thereof, and belongs to the technical field of biology, the protein editing system is successfully constructed on the basis of functional research of RSV M2-1 protein, by utilizing the characteristic that recruited PARN protein is reserved by mutant protein K92A and has an mRNA tail degradation function, and by combining the strong combining capacity of a lambda N-BoxB system, the protein editing system can be used for editing the protein. The system comprises effector plasmids and targeting plasmids, the targeting plasmids aiming at target proteins are edibly constructed, and the targeting plasmids and the effector plasmids are matched to be introduced into host cells, so that the expression of specific target proteins in the cells is remarkably knocked down. The protein editing system can be developed into a product for protein editing, has been successfully applied to knock-down expression of various proteins and pathogens, can avoid irreversible damage of a gene level caused by gene editing, provides a new solution thought for rapid application of gene knock-down expression, and has a wide application prospect.

The invention belongs to the technical field of biological medicines, and particularly relates to application of an alpha-galactosidase inhibitor in preparation of a medicine for treating myocardial injury related diseases. According to the invention, GLA-mediated Gal2Cer / GalCer imbalance found by space metabonomics is taken as a core mechanism, and a precise intervention scheme is provided. The targeting property is strong, the myocardial specificity is prominent, the used gene knock-down vector can focus on myocardial tissues to play a role, and the peripheral side effect risk is reduced; meanwhile, the GLA inhibitor is applied to myocardial I / R injury intervention for the first time, and the GLA inhibitor and gene knock-down synergistically prove that the cardiac function can be effectively protected and pathological injuries such as myocardial infarction and fibrosis can be relieved by regulating GLA, and related effects are verified by animal experiments; a new target is provided for treatment of heart diseases such as myocardial I / R injury, the clinical application scene of the GLA inhibitor is expanded, and the GLA inhibitor has innovation value and clinical transformation potential.

The invention discloses application of inhibition of RUNX3 gene expression in preparation of medicines for preventing or treating silicosis, and belongs to the technical field of biological medicines. In-vivo and in-vitro silicosis models prove that the RUNX3 has high specificity expression in the silicosis process and is a key factor for driving disease progression. By inhibiting the expression of RUNX3 through a gene knock-down technology, silica-induced lung inflammation infiltration and collagenous fiber deposition can be significantly reduced, and the expression levels of key inflammatory factors (IL-1beta, IL-6 and TNF-alpha) and fibrosis indexes (Collagen I, Collagen III and Fibronectin) can be effectively down-regulated. Therefore, any substance capable of specifically inhibiting the expression of the RUNX3 gene, including but not limited to siRNA, shRNA, antisense oligonucleotide, ribozyme, microRNA and an inhibitor targeting the gene promoter thereof, can be used for developing the pharmaceutical composition for preventing or treating silicosis.

The application discloses a use of a DDX51 gene in human melanoma and related products. The application takes the DDX51 gene as a template, designs an RNA interference target sequence, and constructs a target gene RNA interference lentivirus vector. RT-PCR detection of the expression of the target gene in target cells, lentivirus infection of the target cells, qPCR detection of the mRNA level DDX51 gene knockdown efficiency, Celigo detection of the influence of the DDX51 gene knockdown on cell proliferation, detection of the influence of the DDX51 gene knockdown on clone formation, Invasion (Transwell with ECM) detection of the influence of the DDX51 gene knockdown on the cell invasion ability, and the like confirm that the interference target gene DDX51 has the effects of inhibiting the proliferation, clone formation, invasion and metastasis of human melanoma cells A375.

This invention belongs to the field of genetic engineering technology and discloses a shRNA that targets and knocks down the HDAC5 gene. The sense strand of the shRNA is shown in SEQ ID NO.4, and the antisense strand is shown in SEQ ID NO.5. The sense and antisense strands anneal to form a double-stranded DNA with sticky ends. This invention utilizes lentivirus-mediated RNA interference technology to obtain a Vero cell line with stable HDAC5 gene knockdown, demonstrating that HDAC5 gene knockdown can inhibit PPRV proliferation, providing a novel potential target for the development of anti-PPRV drugs.

The invention provides application of ANKRD22 as a target in prognosis evaluation and treatment of cervical cancer. The application of the ANKRD22 as the target refers to reduction of expression of an ANKRD22 gene by adopting gene knockout and gene knockdown. According to the medicine, ANKRD22 is taken as a target, migration and invasion of cervical cancer cells can be remarkably inhibited by knocking down the ANKRD22, and apoptosis and autophagy of the cervical cancer cells are promoted through a PI3K / AKT / mTOR signal channel, so that an anti-tumor effect is achieved in the cervical cancer cells. The invention discloses the effect of ANKRD22 in occurrence and development of cervical cancer and a potential molecular mechanism of the ANKRD22, the ANKRD22 can be used as a potential new target spot for treating cervical cancer, and an ANKRD22 inhibitor is a potential novel medicine for treating cervical cancer.

The invention relates to the technical field of drug delivery, and discloses an MTHFD2siRNA loaded nano-composite as well as a preparation method and application thereof. The nano-composite comprises siRNA and PFeOOH, the siRNA comprises MTHFD2siRNA-1 of a fragment 1 of a target MTHFD2, or MTHFD2siRNA-2 of a fragment 2 of the target MTHFD2, or MTHFD2siRNA-3 of a fragment 3 of the target MTHFD2, and the PFeOOH comprises the siRNA and the PFeOOH. Wherein the sequence of the fragment 1 of the MTHFD2 is as shown in SEQ ID No. 1, and the sequence of the fragment 2 of the MTHFD2 is as shown in SEQ ID No. 2; the sequence of a fragment 3 of the MTHFD2 is as shown in SEQ ID No. 3; the MTHFD2siRNA loaded nano compound provided by the invention can efficiently deliver siRNA, solves the problem of combined delivery of siRNA and an iron agent, and realizes synergistic induction of tumor cell ferroptosis by gene knock-down and iron accumulation.

The application discloses a method for constructing gene editing-free all-male fish based on kdm6bb gene knockdown technology, comprising the following steps: using CRISPR / Cas9 gene editing technology to perform knockdown treatment on fish kdm6bb genes, and obtaining kdm6bb heterozygotes (kdm6bb+ / -) through genotyping; obtaining kdm6bb heterozygous mutants with effectively reduced kdm6bb protein expression through self-crossing of the heterozygote fish; screening pseudo-female fish with XY gender genetic type through gender molecular markers, mating with wild-type XY male fish, screening YY gender genetic type super-male fish without kdm6bb mutation, crossing with wild-type XX female fish, obtaining all-male fish population without carrying kdm6bb mutation, realizing monosex culture of fish, improving the efficiency of gender control breeding, and showing wide adaptability in aquatic economic fish based on XX / XY gender determination.

The invention provides application of an MDH2 inhibitor to treatment of abnormal glucose metabolism diseases and preparation of drugs for treating abnormal glucose metabolism. Through gene knockdown or knockout and drug intervention, it is found that MDH2 plays a key role in central glycometabolism induction, and targeting MDH2 may provide a new treatment method for improving the blood glucose level of diabetic patients. The application also finds that the LW6 can improve glucose metabolism disorder and treat diabetes mellitus by targeting an MDH2 target, verifies the effectiveness and safety of the LW6 under central and peripheral administration conditions, does not cause weight loss, eating reduction or other metabolism-related side effects, and reveals a potential neurological mechanism and clinical application value of the LW6.