181 results about "Small RNA" patented technology

The invention relates to a biological preparation for improving quality of weever, the biological preparation comprises an injection and an edible agent, the injection comprises chemically modified chemically synthesized single-stranded small RNA, Rev-erbalpha protein and Per1 protein, and the edible agent comprises Clock core peptide 1, Clock core peptide 2 and taurine. Skeletal muscle development, liver lipid metabolism and generation of cell inflammatory factors of the weever are influenced by adding and injecting the feed, so that the quality of the weever is improved, intramuscular fat deposition is promoted, the taste and flavor of fish meat are improved, the energy reserve of the weever can be improved, and the culture cycle is shortened. The improvement of cell inflammatory factors shows that the disease resistance of the weever is improved, the survival rate in the breeding process is further improved, and the use amount of antibiotics is reduced. The method provides an important theoretical basis for improving fish meat quality in China and promoting high-quality, efficient and healthy development of aquaculture, and has a wide market prospect.

The invention discloses a novel tsRNA high-sensitivity detection kit based on CRISPR (clustered regularly interspaced short palindromic repeats) / Cas13a-Cas14a, and belongs to the field of tumor marker detection. The tsRNA (tRNA-derived small RNA) is a diagnostic marker which is found recently and has great potential. In order to realize the clinical transformation of tsRNA as soon as possible, the tsRNA detection kit is developed on the basis of nuclease P1 combined with CRISPR / Cas13a-Cas14a linkage dual systems, and comprehensive, ultra-sensitive, high-specificity, low-cost and rapid tsRNA detection is realized through multiple signal amplification. Compared with the traditional technology, the sensitivity of the method is improved by 105-107 times, 1% of tsRNA in the mixture can be detected, and the method has more advantages in clinical application. Therefore, a new scheme is provided for application of the tsRNA in clinical diagnosis of diseases, and meanwhile, a reliable technology is provided for mechanism research of the tsRNA in occurrence and development of the diseases.

The invention belongs to the field of biological detection, and discloses an exosome RNA detection composition based on positive and negative targets and a use method thereof, the composition comprises two positive targets and one negative target: the positive target A is miRNA stably expressed in exosome and is used for reflecting the expression level of small RNA; the positive target B is typical mRNA and is used for evaluating the quality of the long-fragment RNA; the negative target C is bacterial 16S rRNA and is used for identifying bacterial nucleic acid pollution in a sample. The method combines III-generation reverse transcriptase and HotStart Taq enzyme to construct a qPCR system, has high sensitivity and specificity, and is suitable for rapid detection of trace RNA. Whether the exosome RNA is suitable for downstream analysis or not can be judged by acquiring the Cq values of the three types of targets and setting a reference range. According to the invention, the integration of quality control, quantification and pollution identification of the exosome RNA is realized.

The invention relates to a method and application for positively regulating the defense response of plants to pests, and the method comprises the following steps: the plants are contacted with small RNA (Ribonucleic Acid) to enhance the resistance of the plants to the pests, and the small RNA comprises plant endogenous 22nt-siRNA. Researches find that the endogenous 22nt-siRNA of the plant can improve the resistance of the plant to phytophagous insects, and the 22nt-siRNA can specifically target a field phytophagous insect gene while enhancing the resistance of the plant, has the biological prevention and control advantage of no environmental pollution, and can be used for preventing and treating the phytophagous insects. Therefore, the method plays an important role in forward regulation of defense response of plants to pests and prevention and control of plant pests.

The invention belongs to the field of meat pigeon breeding, and particularly relates to miRNAs capable of serving as a trichomonad pigeon biomarker and application of the miRNAs. The invention provides application of miRNAs as a trichomonas pigeonae biomarker in preparation of a trichomonas pigeonae diagnostic reagent. According to the invention, high-throughput sequencing and bioinformatics analysis are simultaneously carried out on oral mucosa small RNA and transcriptome of a trichomonad infected positive individual and a healthy individual for the first time, so that an epigenetic molecular mechanism of oral mucosa tissue of a trichomonad infected regulation body is explained by mining a key significant difference miRNA-mRNA network.

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 invention provides application of plasmodiophora brassicae infected early-stage chitin in prevention and control of cruciferae clubroot, and relates to a technology and application for preventing and controlling the cruciferae clubroot based on blocking synthesis of zoosporangium chitin in plasmodiophora brassicae infected early-stage chitin. Experiments prove that chitin components exist in zoosporangia in the early stage of plasmodiophora brassicae infection and are synthesized by plasmodiophora brassicae chitin synthetase genes PbCHS3 and PbCHS7, and a new target is provided for prevention and control of clubroot of cruciferae crops. The clubroot can be efficiently prevented and controlled by expressing PbCHS3 and PbCHS7 silencing vectors in arabidopsis thaliana and oilseed rape and treating or applying pesticide polyoxin capable of blocking chitin synthesis through exogenous small RNA, and a key technology and a key method are provided for preventing and controlling the clubroot of cruciferae.

The invention belongs to the technical field of gene engineering, and particularly relates to application of corn zma-miR319 and / or a target gene of the corn zma-miR319 in regulation and control of corn stem rot resistance. According to the application, sequencing data of small RNA, a degradation group and a transcriptome are integrated, a miRNA family member zma-miR319 is identified, and the resistance of corn to stem rot can be positively regulated and enhanced; the target gene ZmMYB74 is a core regulation gene of an Fg response module, the stem rot resistance of a transgenic plant over-expressing the ZmMYB74 is weakened, and lignin deposition is reduced; the resistance is obviously enhanced by knocking out or inhibiting ZmMYB74, the ZmMYB74 is used as a transcription inhibition factor, the expression of a lignin synthesis related gene ZmCAD is negatively regulated and controlled, and the separation of the disease-resistant gene ZmMYB74 not only contributes to the cultivation of a persistent disease-resistant corn variety, but also deepens the understanding of a stem rot resistance molecular mechanism.

The present invention refers to a method for isolating a fraction enriched of small RNA molecules from a fungal or plant sample.

The invention relates to an ultramicro small RNA (Ribonucleic Acid) sequencing method, which adopts a tubular reaction, carries out RNA terminal repair and AlKB demodification enzyme activity reaction by expanding a system step by step, and comprises cell lysis, RNA terminal repair, 3'adapter connection, AlKB demodification enzyme activity reaction, RNA purification, RT (Reverse Transcription) annealing, 5 'adapter connection, cDNA (Complementary Deoxyribose Nucleic Acid) reverse transcription, library amplification and NGS (Next Generation System). According to the present invention, the improvement is performed on the basis of the existing PANDORA-Seq technology, the efficient and accurate detection of the trace cell small RNA is achieved by optimizing the reaction conditions, and the UltraTra-PANDORA-Seq method is successfully established. According to the method, the technical problem of the traditional method in trace cell small RNA detection is solved, and a feasible and reliable analysis means is provided for deep research on the functional mechanism of the small RNA in a living body. The development of the Ultra-PANDORA-Seq technology provides important scientific basis and technical support for exploration of intergenerational genetic mechanisms of metabolic diseases, early clinical diagnosis, development of small RNA drugs and clinical intervention.

The invention discloses a novel tRNA (transfer ribonucleic acid) derived small RNA biomarker tRF5-23-GlyTCC-2 and application thereof, and relates to the technical field of biological medicines. The nucleotide sequence of the biomarker tRF5-23-GlyTCC-2 disclosed by the invention is as shown in SEQ ID NO. 1, and the nucleotide sequence of the biomarker tRF5-23-GlyTCC-2 is as shown in SEQ ID NO. Results of embodiments show that the gene has significant low expression in colorectal cancer tissues and serum, when the gene is independently used for colorectal cancer diagnosis, AUC for distinguishing a patient from a healthy control reaches 0.8628, the sensitivity is 74%, the specificity is 92%, and the AUC, the sensitivity and the specificity are both superior to CEA; the AUC is improved to 0.9077 and the sensitivity reaches 90% when the compound is used in combination with CEA (Carcino Embryonic Acid). Meanwhile, low expression of the marker is related to poor prognosis characteristics such as late T staging and low differentiation of a patient, and the marker can be used for prognosis evaluation. Functional experiments prove that overexpression can inhibit colorectal cancer cell proliferation and tumor growth, and a new scheme is provided for colorectal cancer diagnosis and treatment.

The application discloses two kinds of small RNA compositions for improving biological control effect of termites, and microRNA-like RNA (milRNA) sequences are milR-32 and milR-48 respectively. Double-stranded RNA can enhance the effect of the beetle green fungus on killing termites. The milR-32 and the milR-48 belong to the milRNA coded by the green fungus, and the gene sequences are derived from the transcriptome database of the beetle green fungus. The milR-32 and the milR-48 mimics are designed and synthesized based on the milR-32 and the milR-48 sequences respectively. After the beetle green fungus is used to infect the termites taking the milR-32 or the milR-48 mimic, the infection mortality of the termites is significantly increased. The mimic designed based on the milR-32 and the milR-48 sequences can significantly improve the biological control effect of the termites.

The present application discloses a sequencing method, sequencing reagent and application for Small RNA. The present application is a sequencing method for Small RNA, comprising adding a normal sequencing primer and a blocking sequencing primer to a primer solution when sequencing a circular library of a mixed sample, and using a mixed primer for sequencing; the blocking sequencing primer is formed by chemically modifying the 3' terminal hydroxyl group of the normal sequencing primer. The sequencing method of the present application, by adding a blocking sequencing primer, enables Small RNA sequencing of a circular library of a mixed sample, and can effectively ensure the sequencing of the label sequence, thereby improving the data utilization rate and obtaining a good resolution rate. The Small RNA sequencing method and sequencing reagent of the present application provide a new idea and solution for Small RNA mixed sample sequencing, solving the current problem of sequencing resource waste in Small RNA sequencing of circular libraries.

The invention provides application of a ring finger protein coding gene OsRF in regulation and control of rice grain shape and grain weight, the gene OsRF for coding the ring finger protein is separated and cloned from rice Nipponbare, and the gene is targeted by rice small RNA OsmiR530. In order to prevent the OsmiR530 from being cut and improve the expression level of the OsmiR530, artificial synonymous mutation is carried out on the OsRF recognized sequence by the OsmiR530, a mutant OsRF fragment (mOsRF) of which the recognition sequence is subjected to synonymous mutation is created by utilizing overlapping PCR (Polymerase Chain Reaction), an overexpression vector of the mOsRF is constructed and is overexpressed in rice, and the grain width and the grain weight of transgenic rice are obviously increased. Therefore, the overexpression of the mOsRF in the rice is of great significance for obtaining round-grain rice and increasing the rice yield, and a new resource and a new research method are provided for rice grain shape improvement breeding.

The present disclosure relates generally to the use of small RNA biomarkers in a biological sample for detection of residual cancer cells in a patient who has received treatment for a cancer.

The invention provides a method for removing a linker self-linking product in a small RNA library preparation process. The method comprises the following steps: step a) adding excessive reverse transcription primers to close a 3'linker; a step b) of Damp; l, enzyme treatment; and a step c) of Cas-sgRNA complex treatment. The invention also provides a method for removing a linker self-linking product and polluted nucleic acid in the preparation process of the small RNA library, a preparation method of the small RNA library, a multi-omics sequencing library containing the small RNA and the like.

Provided is an improved design of shRNA based on structural mimics of miR-451 precursors. These miR-451 shRNA mimics are channeled through a novel small RNA biogenesis pathway, require AGO2 catalysis and are processed by Drosha but are independent of DICER processing. This miRNA pathway feeds active elements only into Ago2 because of its unique catalytic activity. These data demonstrate that this newly identified small RNA biogenesis pathway can be exploited in vivo to produce active molecules.

The invention provides a variable cleavage polypeptide and application thereof, the variable cleavage polypeptide is a polypeptide composed of 23 amino acids at the N terminal of calendula spica HBFD, and can realize specific cleavage in tobacco and multi-protein tandem expression of a single expression cassette. The cleavage specificity of the variable cleavage polypeptide in plants is superior to that of a previously reported small RNA virus 2A self-cleavage polypeptide, and reduction of non-specific cleavage is facilitated. The variable cleavage polypeptide is a protein cleavage element derived from a plant, and the application value of a multi-protein tandem expression technology based on a single expression cassette in a plant bioreactor is improved.

This disclosure relates to the field of nucleic acid drug technologies, and in particular to a small RNA-based drug, preparation, and use thereof in prophylaxis and / or treatment of cardiomyopathy. Provided is a novel small RNA with the nucleotide sequence of SEQ ID NO: 10. The small RNA exhibits good structural stability, marked efficacy at 24 h, and high affinity to nascent polypeptide-associated complex α (NACα) protein. With strong activity to suppress the expression of pyroptosis-related proteins, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and gasdermin D (GSDMD), the small RNA can effectively protect cardiomyocytes and enhance the therapeutic effect on diabetic cardiomyopathy (DCM). Furthermore, the small RNA posses advantages of low effective concentration and high biosafety, highlighting its potential for clinical drug development and application as a nucleic acid drug.

The invention belongs to the technical field of RNA modification detection, and particularly relates to a method for distinguishing four terminal small RNAs through APB small RNA imprinting hybridization. RNA to be detected is subjected to phosphatase and T4 polynucleotide kinase treatment, RNA purification and APB small RNA imprinting hybridization, the response condition of the RNA to phosphatase and T4 polynucleotide kinase is obtained by comparing whether the electrophoresis migration rate before and after treatment is changed or not, and then the terminal types of 2 ', 3' OH, 2 'OMe, 2', 3'cP and 3 'P before treatment are deduced. Compared with a traditional small RNA imprinting hybridization method, the method has the advantages that the migration rate difference of bands of different modified small RNAs during APB gel electrophoresis is larger, and the problems that the migration rate difference of different modified RNAs is small, and experimental results are often difficult to judge due to the influence of RNA purity, concentration and experimental conditions in a traditional method are solved; the method has a good application prospect in the fields of small RNA modification and functional research thereof.

According to the invention, reverse transcriptase Marathon or a mutant thereof is fused with Cas nuclease, a small RNA binding exonuclease protection factor La and a positioning signal peptide to form a fusion protein capable of guiding editing. Experiments show that a guide editing system based on the fusion protein can realize base replacement operation in rice callus, and has important application value in the field of biological breeding.

The present invention provides a serum exosomal tsRNA (tRNA-derived small RNA, tsRNA) biomarker, a probe and their applications for the diagnosis of lupus nephritis, belonging to the field of molecular biology; the biomarker is one or a combination of tRF-Thr-TGT-4-M3 and tRF-Tyr-GTA-1-M2; the exosomal tsRNA screened by the present invention is significantly highly expressed in the serum exosomes of lupus nephritis patients, has a good correlation with the clinical indicators of lupus nephritis, and can be used to identify whether SLE patients have kidney involvement; the biomarker can be used to prepare a diagnostic kit for the auxiliary early diagnosis of lupus nephritis in clinical practice.

The present disclosure relates generally to the use of small RNA biomarkers in a biological sample for early detection of invasive breast cancer (IBC) and noninvasive ductal carcinoma in-situ (DCIS).

The application belongs to the technical field of nucleic acid drugs, and particularly relates to a small fragment RNA derived from circular RNA and application of the small fragment RNA in treatment of diabetic vascular diseases. The nucleotide sequence of the small fragment RNA is CUAGCCGAAUGUUAAAAAAU. The small fragment RNA provided by the application has the effect of protecting and maintaining the biological function of vascular endothelial cells, has good structural stability, has a long half-life in the body, has the advantages of low effective concentration and high biological safety, has good application potential in the field of clinical nucleic acid drug development, and provides a new idea for treatment of chronic and progressive vascular lesions caused by diabetes.

The present invention relates to a resistance-free plasmid production system. The present invention combines toxic transcripts / anti-toxic transcripts, operon-related elements and small RNAs to construct a plasmid comprising a nucleic acid fragment 3 encoding a small RNA, and host cell expression cassettes comprising a toxic gene expression cassette and an anti-toxic gene expression cassette, wherein the toxic gene expression cassette comprises a promoter and a nucleic acid fragment 1 for transcribing the toxic transcript, and the anti-toxic gene expression cassette comprises a promoter, an operon and a nucleic acid fragment 2 for transcribing the anti-toxic transcript. The reverse complementation of the nucleic acid fragment 3 and the nucleic acid fragment 1 produces a steric effect to inhibit the function of the toxic transcript, so that the entire plasmid production process does not need to be added with antibiotics or other inhibitors to realize the screening and production of the plasmid, thereby the harm of antibiotics and inhibitors to the plasmid in cell gene therapy is solved.

The invention discloses a multi-omics plant phenotype prediction method fused with small RNA, and relates to the field of bioinformatics and agricultural biology, and the method comprises the following steps: obtaining and preprocessing genotype, transcript expression and small RNA expression data to obtain a standardized feature matrix; performing feature screening based on Pearson correlation on the standardized feature matrix to obtain corresponding genotype, transcript expression and small RNA expression feature subsets; and based on a pre-trained MirGP prediction model, through multi-branch feature extraction, multistage efficient channel attention fusion and regression prediction, outputting a plant phenotype prediction result. According to the method, multiple types of small RNA regulation and control features are introduced and a hierarchical deep learning fusion architecture is constructed, so that multiple omics feature dimensions are expanded, data redundancy is reduced, and effective integration and deep mining of cross-modal features are realized.

The application belongs to the field of pharmaceutical chemical industry and relates to a cyclic peptide compound shown as formula M or a stereoisomer, a tautomer or a mixture thereof of the compound, a pharmaceutically acceptable salt, a polymorph, a co-crystal or a solvate of the compound, or a stable isotope derivative, a metabolite or a prodrug of the compound. The application also relates to a method for synthesizing the cyclic peptide compound and an intermediate. The provided cyclic peptide compound has a strong inhibitory effect on a coronavirus 3CL protease and a small RNA virus (for example, an enterovirus EV71 virus) 3C protease. After cell level activity testing, the provided cyclic peptide compound has an obvious inhibitory effect on EV71 virus and coronavirus and the like, indicating that the cyclic peptide compound shown as formula M can produce a viral inhibitory effect at the cell level by inhibiting viral protease activity, and has a good application prospect as an antiviral drug.

A composition for treating cancer is disclosed. The composition includes a lentiviral particle and an aminobisphosphonate drug. The lentiviral particle is capable of infecting a target cell, such as a cancer cell, and includes an envelope protein optimized for targeting such target cell and a viral vector. The viral vector includes a small RNA optimized to target an FDPS mRNA sequence. The aminobisphosphonate drug includes zoledronic acid.

The present invention relates to a method classifying target RNA modifications in a neural network system, to a method for training a neural network system that is configured to classify target RNA modifications, to a neural network system, to a computer system adapted to classify target RNA modifications, to a computer program and to a non-transitory readable medium.