212 results about "Gene silencing" patented technology

The invention discloses three pattern recognition protein Hc beta GRP genes for preventing and treating fall webworms, dsRNA and application thereof. Specifically, the nucleotide sequences of the three pattern recognition protein Hc beta GRP genes for preventing and treating fall webworms are respectively SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3. The recombinant Hc beta GRP protein is successfully obtained through a prokaryotic expression system, an in-vitro bacteriostatic test proves that the recombinant Hc beta GRP protein, the Hc beta GRP protein and the dsRNA have direct bacteriostatic activity, dsRNA is designed based on a pattern recognition protein Hc beta GRP gene, a hyphantria cunea Hc beta GRP dsRNA interference system implemented by using an injection method is established, an obvious gene silencing effect is obtained by injecting the Hc beta GRP dsRNA interference system into hyphantria cunea, and the Hc beta GRP dsRNA interference system is applied to the hyphantria cunea. The technical support is provided for the application of the Hc beta GRP RNAi technology. Besides, the dsRNA of the hyphantria cunea Hc beta GRP is combined with the biocontrol bacteria by utilizing an established hyphantria cunea Hc beta GRP RNA interference system, so that the control effect of the biocontrol bacteria can be remarkably improved.

The invention provides an hpRNA interference vector for laver and application of the hpRNA interference vector, and belongs to the technical field of genetic engineering and algae biology. According to the hpRNA interference vector, a binary vector pBI121 is used as a skeleton vector, a laver endogenous Actin promoter and a multiple cloning site segment are inserted, the multiple cloning sites allow target gene segments to be cloned in a forward and reverse repetition mode, the target gene segments are spaced by introns, and therefore an efficient hpRNA structure is formed after transcription, and the efficient hpRNA interference vector is obtained. The gene can be directly used for silencing important genes related to growth and development, metabolic regulation, stress response and the like in the laver. The invention further provides a construction method of the hpRNA interference vector and application of the hpRNA interference vector in silencing laver genes and provides a method for silencing laver gene expression, the gene silencing effect is remarkable, experimental results can be repeated, and a powerful tool is provided for molecular breeding and genetic improvement of laver.

The invention discloses an upland cotton GhCRK29 gene as well as an encoding protein and application thereof, the GhCRK29 gene is separated from cotton, the nucleotide sequence of the GhCRK29 gene is as shown in SEQ ID No.1, and the amino acid sequence of the encoding protein of the GhCRK29 gene is as shown in SEQ ID No.2. According to the invention, a virus-induced gene silencing technology and a transgenic overexpression technology are further utilized to carry out preliminary research on the function of the cotton GhCRK29 gene in cotton verticillium wilt resistance, and a result shows that the verticillium wilt resistance of a cotton seedling plant is obviously reduced after the GhCRK29 gene in cotton is silenced; after the GhCRK29 gene is over-expressed in arabidopsis thaliana or cotton, the verticillium wilt resistance of arabidopsis thaliana or cotton is remarkably improved, and the resistance of a GhCRK29 gene forward regulation plant to verticillium dahliae is determined; the invention has application prospects in the aspects of improving the verticillium wilt resistance of plants or cultivating verticillium wilt resistant plant varieties and the like.

The invention discloses a verticillium dahliae virulence gene, a verticillium dahliae virulence protein and application. The invention relates to the technical field of biology, and provides a method for preventing and treating cotton verticillium wilt based on RNAi (RNA interference) as well as related components and application thereof. Specifically, the invention discloses application of the verticillium wilt prevention and treatment by a method for inhibiting or silencing the expression of verticillium dahliae pathogenic gene VdHDAC (coding protein of the verticillium dahliae pathogenic gene VdHDAC is shown as SEQ ID NO: 3), the core of the verticillium wihliae pathogenic gene VdHDAC is to provide dsRNA molecules capable of targeting the gene, and the positive-sense strand of the dsRNA molecules is preferably selected from SEQ ID NO: 5, SEQ ID NO: 8, SEQ ID NO: 9 or SEQ ID NO: 11. Based on the molecule, the invention further provides a recombinant vector containing the coding sequence, a recombinant microorganism, a transgenic disease-resistant plant and an RNAi pesticide preparation. The scheme can be realized through host-induced gene silencing, microorganism-mediated gene silencing or preparation treatment and the like, the virulence of pathogenic bacteria can be effectively reduced, and a new technical approach is provided for green prevention and control of verticillium wilt.

The invention relates to the technical field of plant genetic engineering, and particularly provides cloning of a GbGELP25D gene of gossypium barbadense and application of the GbGELP25D gene of gossypium barbadense. According to the invention, the GbGELP25D gene with a full length of 1092 bp is cloned from the sea island cotton variety Xinhai No. 7 for the first time, and the gene encodes a secretory lipase protein with a signal peptide and is positioned in an extracellular gap. Through bioinformatics analysis, phylogenetic classification, protein structure modeling and signal peptide function verification, it is clear that the gene belongs to a plant GELP family. Furthermore, a virus-induced gene silencing technology is utilized to prove that the silent GbGELP25D can obviously enhance the resistance of cotton to verticillium wilt, and the mechanism of the silent GbGELP25D is closely related to activation of ethylene synthesis and signal channels and induction of expression of disease-resistant related genes. The resistance gene provided by the invention enriches gene resources of cotton verticillium wilt resistance breeding, and has important theoretical significance and application value.

This invention belongs to the field of plant genetic engineering technology, specifically relating to the application of H3K27me3 in regulating the cotton response to potassium chloride stress. By integrating CUT & Tag chromatin analysis and RNA-seq, this invention demonstrates that potassium chloride stress induces a reduction in H3K27me3 deposition across the entire genome, accompanied by characteristic stress phenotypes in cotton seedlings. Inhibition of H3K27me3 using RDS 3434 significantly improved KCl-induced physiological damage, confirming the functional correlation between this epigenetic marker and stress tolerance. Furthermore, virus-induced gene silencing confirmed that genes associated with H3K27me3 are important components of the cotton ion stress response network. Therefore, this invention elucidates the epigenetic landscape regulating adaptation to potassium chloride stress.

The invention discloses an inflorescence gene silencing method which comprises the following steps: firstly, synthesizing a specific nucleotide fragment according to a target gene, and constructing the specific nucleotide fragment to a plant virus vector; introducing the constructed plant virus vector and the corresponding helper plasmid into the inflorescence; soaking the inflorescence in a metal chelating agent salt solution; and finally, screening to obtain plants with inflorescences subjected to target gene silencing. The chrysanthemum capitulum is selected as an infection object, the silencing method is optimized, the target gene silencing efficiency is remarkably improved, the phenotype detection period after infection is remarkably shortened, and the method is efficient, convenient and high in operability. Meanwhile, the method can effectively avoid the cross protection phenomenon of plants, overcomes the obstacle of limited transmission range of viral vectors, does not need to consider the problems of death and growth cycle of tissue culture seedlings or seedlings after infection, and provides a feasible means for rapid identification of chrysanthemum gene functions and large-scale development of chrysanthemum gene function research.

The invention discloses a nucleic acid construct for inducing DDB2 gene silencing and application of the nucleic acid construct. The designed targeted expression system based on the adeno-associated virus (AAV) vector is higher in specificity, non-target tissue can be prevented from being interfered, the tissue specificity of gene intervention is improved, and the off-target risk is small; the shRNA has high stability and processing efficiency, reduces immune response and interference effect, has high expression efficiency and strong histocompatibility, and is suitable for long-term expression intervention in vivo.

An engineered transcriptional modulator (ETM) comprising: (a) at least one epigenetic effector domain; operably linked to (b) an endonuclease.

The application provides application of a wheat ELP2 gene in regulating resistance of wheat to stripe rust, and belongs to the technical field of genetic engineering. The application provides application of the wheat ELP2 gene in regulating resistance of wheat to stripe rust, also provides a silencing fragment of the wheat ELP2 gene obtained by an amplification method, and gene silencing of the wheat ELP2 gene is carried out by using the silencing fragment, and in the examples, overexpression plants of the ELP2 gene are constructed, it is found that the overexpression weakens the resistance of wheat to non-compatible races of Puccinia striiformis f. sp. tritici, the interaction relationship between ELP2 and the PAZ motif of DCL1 is verified by using yeast double hybridization, LCI and Pull-down, meanwhile, the resistance of wheat to compatible races is enhanced and the infection of Puccinia striiformis f. sp. tritici is inhibited after silencing DCL1 in wheat by VIGS, therefore, the ELP2 gene plays a disease function in the interaction between wheat and Puccinia striiformis f. sp. tritici.

The invention discloses a transcription factor TgWRKY65 of torreya grandis as well as a coding gene and application of the transcription factor TgWRKY65. The amino acid sequence of the transcription factor TgWRKY65 is shown as SEQ ID NO.2, and the nucleotide sequence of the coding gene is shown as SEQ ID NO.1. The invention further discloses a preparation method of the transcription factor TgWRKY65. According to the invention, a homologous expression material is constructed through cloning of the TgWRKY65 gene, a transgenic technology and gene silencing, and the application prospect in the aspect of increasing the content of the torreya grandis catechin is great; when the coding gene is over-expressed, the content of catechin is increased; when the coding gene is silenced, the content of catechin is reduced.

Disclosed are a poly-oligonucleotide capable of triggering gene silencing and a use thereof, and specifically disclosed are a compound represented by formula (I) and the use thereof.

This invention discloses a chimeric guide RNA integrating RNA interference and CRISPR-Cas13d and its applications. Utilizing the structural matching between shRNA and Cas13d crRNA, this invention embeds the complete Cas13d crRNA into the classic shRNA backbone, constructing a dual-pathway synergistic silencing chimeric guide RNA (SS-Rx). Experiments have demonstrated that SS-Rx possesses the dual activity of shRNA and CRISPR-Cas13d, exhibiting superior gene silencing efficiency compared to traditional RNA silencing tools, and can inhibit the proliferation, migration, and invasion of bladder cancer cells by suppressing tumor-related genes. This invention synergistically combines the advantages of RNA interference and CRISPR-Cas13d, providing an efficient and widely applicable framework for gene function research and RNA-targeted therapy, with significant economic benefits and broad application prospects.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a PROSiCAPE14 gene and a protein encoded by the PROSiCAPE14 gene to improvement of plant salt tolerance. According to the invention, it is found for the first time that salt stress can cause down-regulation of the expression of the millet PROSiCAPE14 gene. Under a salt treatment condition, the plant height and the root length of a PROSiCAPE14 gene coded small peptide externally applied to millet and a constructed PROSiCAPE14 gene overexpression strain are obviously worse than those of a control group, and the plant height and the root length of a PROSiCAPE14 gene silent plant are obviously better than those of the control group. It is indicated that the PROSiCAPE14 gene plays an important role in the salt stress response process of the millet, and a foundation is provided for further research on the effect of the CAPE14 gene in the adaptation of the millet to abiotic stress and other possible biological functions.

The application discloses a protein kinase gene positively regulating drought resistance of oats AVESA.00022b.r1.6C0001455 and application thereof, and belongs to the technical field of plant genetic engineering. AVESA.00022b.r1.6C0001455 The CDS sequence of the gene is shown as SEQ ID NO. 1, and the amino acid sequence is shown as SEQ ID NO. 2. Expression mode analysis shows that the gene is significantly induced and up-regulated under drought stress; virus-induced gene silencing verification shows that silencing the gene can significantly reduce the drought resistance of the oats; overexpression verification shows that overexpression of the gene can significantly enhance the drought resistance of the oats, and the performance is that wilting is delayed, the water loss rate is reduced, and the survival rate is improved. The application first proves that the gene is a key positive regulation factor of drought resistance in the oats, and can be used for cultivating drought-resistant transgenic plants and as a drought-resistant molecular marker for assisted breeding, and has important application value in genetic improvement of crop drought resistance.

The invention discloses a LeRho1 gene expression vector and a method for improving heat resistance and trichoderma resistance of lentinus edodes. The LeRho1 gene expression vector is a recombinant expression vector, a target gene for coding the LeRho1 protein is inserted into the recombinant expression vector, and the amino acid sequence of the LeRho1 protein is shown as SEQ ID NO. 1. The invention finds that the LeRho1 gene has a positive regulation effect on the heat resistance of the shiitake mushroom strain, and the LeUSPA and LeUSPH genes mediate the thermal response of the shiitake mushroom strain through negative regulation. Therefore, according to the method, the LeRho1 gene overexpression vector or LeUSPA and LeUSPH gene silencing vectors are constructed and transferred into the shiitake mushroom strain for genetic transformation, the heat resistance and trichoderma resistance of the shiitake mushroom strain can be remarkably improved, and the transformation efficiency is high.

The invention belongs to the technical field of gene intervention, and provides a feeding-mediated marine mussel gene silencing method which comprises the following steps: linearizing plasmids to obtain linearized plasmids; determining a cDNA fragment of the target gene from the target mussel, and performing amplification by using the specific primer pair to obtain a target gene fragment; connecting the target gene segment with the linearized plasmid to obtain a connection product; transforming the connection product into escherichia coli, screening to obtain a positive strain, extracting target recombinant plasmids, and transforming the target recombinant plasmids into escherichia coli for expressing dsRNA to obtain engineering bacteria; engineering bacteria and microalgae are mixed according to the cell number ratio of (90-110): (1-2), a bacteria-algae feeding mixture is obtained, and mussels are fed with artificial seawater containing the bacteria-algae feeding mixture for 1-10 d. The method disclosed by the invention does not cause antioxidant system disorder or energy metabolism abnormality, and is a gene silencing strategy with higher biological safety.

The invention provides a synergistic treatment pharmaceutical composition for preventing and treating recurrence after hepatocellular carcinoma thermal ablation and application of the synergistic treatment pharmaceutical composition. Aiming at malignant tumor progression driven by ACSS3 gene silencing after thermal ablation, an epigenetic editing technology is innovatively combined with metabolite supplementation, and a preparation for specifically activating or promoting ACSS3 gene expression and sodium propionate are cooperatively used, so that ACSS3 can be remarkably reactivated or expression can be promoted in a xenograft model derived from a cell line and a patient; the compound can be used for recovering propionyl-coenzyme A synthesis and normal propionic acid metabolism and inhibiting fatty acid beta-oxidation, so that the growth and metastasis of post-ablation hepatocellular carcinoma (HCC) are effectively blocked, and the limitation of a single therapy is overcome. A clearer mechanism target is provided for preventing recurrence after HCC thermal ablation, meanwhile, the prepared targeting nano-liposome preparation has a synergistic effect, and as a brand-new treatment strategy, the defect that intervention on recurrence roots is insufficient in the prior art is overcome.

The invention relates to the technical field of biology, in particular to a vitis davidii susceptible gene VdTLP19 as well as an encoding protein and application of the vitis davidii susceptible gene VdTLP19. The vitis davidii susceptible gene VdTLP19 is located on a chromosome 2 of vitis davidii and is distributed in a region 3655997-3656642, and the nucleotide sequence of the vitis davidii susceptible gene VdTLP19 is shown as SEQ ID No.1. The amino acid sequence of the protein coded by the gene is as shown in SEQ ID No.2. And after the vitis davidii susceptible gene VdTLP19 is silenced, the resistance of grape fruits to pathogenic bacteria is enhanced. After the VdTLP19 gene provided by the invention is silenced, the resistance of grapes to anthracnose can be improved, and a theoretical basis is provided for grape anthracnose resistance breeding.

The invention discloses application of a flammulina velutipes transcription factor gene AreA in regulation and control of cell wall polysaccharide synthesis and sporocarp taste. Flammulina velutipes as an important edible mushroom is widely welcomed due to rich nutrition, but the edible experience of consumers is seriously influenced and the market potential of the flammulina velutipes is limited due to the problems that the flammulina velutipes is poor in chewiness, easy to plug teeth and the like. The silent strain of the enoki mushroom areA gene is successfully constructed through an RNA interference technology, it is found that the polysaccharide content of cell walls and the toughness of sporocarp can be reduced through reduction of gene expression, then the taste of the sporocarp is improved, and the market competitiveness of the sporocarp is improved. According to the invention, the areA gene in the flammulina velutipes is effectively silenced through the specifically designed siRNA fragment, so that the polysaccharide content of the cell wall and the toughness of the sporocarp are reduced, and a theoretical basis is provided for reducing the toughness of the cell wall of the flammulina velutipes by using a genetic engineering technical means.

This invention relates to plant genetic engineering, and more particularly to a variegated Phalaenopsis orchid and its preparation method, the preparation method comprising the following steps: (1) according to the target gene with the base sequence shown in SEQ ID No. 1 PePDS (1) Synthesize specific nucleotide fragments; (2) Construct specific nucleotide fragments into a plant virus vector; (3) Introduce the recombinant plant virus vector obtained in step (2) into Phalaenopsis orchid plant material; (4) Screen to obtain Phalaenopsis orchids with silenced target genes. This invention achieves rapid, efficient, and non-transgenic leaf color variegation creation in commercial Phalaenopsis orchid varieties for the first time through VIGS technology; it not only breaks through the technical bottleneck of difficult and long cycle of genetic manipulation of orchid plants, significantly shortening the trait verification cycle, but also induces diverse and highly ornamental variegated leaf phenotypes such as albino, yellow, or purple in different varieties, and has the advantages of simple operation, low cost, and industrialization feasibility.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a related biological product of an LhHB4 gene in regulation and control of accumulation of anthocyanin in plants. The nucleotide sequence of the LhHB4 gene is as shown in SEQ ID NO. 1, and the sequence of the encoded protein is as shown in SEQ ID NO. 2. Experiments show that the expression of the LhHB4 is in negative correlation with the biosynthesis of the anthocyanin of the lily, the accumulation of the anthocyanin in the lilies in the lilies can be remarkably inhibited by the instantaneous overexpression of the LhHB4 in the lilies, the accumulation of the anthocyanin in the lilies in the lilies in the leaves is promoted after the LhHB4 gene is silenced, and the regulation of the LhHB4 on the biosynthesis of the anthocyanin depends on an EAR motif. The related biological product of the LhHB4 gene provided by the invention provides important application support for cultivating new varieties with peculiar colors of lilies and improving the ornamental and commercial values of the lilies.

The invention belongs to the field of biotechnology and reproductive medicine, and relates to an application of a cytoskeletal regulatory factor ROCK2 in remodeling male reproductive hormone synthesis, the application is to prepare a preparation for regulating male reproductive hormone synthesis, the preparation is a gene silencing preparation, the active component of the preparation is a small interfering RNA molecule aiming at ROCK2, and the active component of the preparation is a small interfering RNA molecule aiming at ROCK2. By inhibiting expression and activity of ROCK2 in a target cell, enrichment of a cortex F-actin network in a cell cortex and formation of a ring structure are promoted, the level of a phosphorylated myosin light chain is up-regulated, and then the targeted transport efficiency of cholesterol in the cell to mitochondria is enhanced. According to the present invention, the structure is complete, the function is complete, the skeleton stability and the hormone synthesis function of the Redis cell are effectively maintained through the specific inhibition of the ROCK2 expression, the technical threshold and the time cost of the drug screening and evaluation are reduced through the established standardized detection system, and the controllability and the safety of the hormone regulation process are ensured.

The invention relates to a method for regulating and controlling a flower type by silencing a peony organ gene based on a transient expression system, which belongs to a peony flower type regulation and control method and comprises the following steps of: designing a specific primer aiming at a target gene PsuAG, obtaining a target fragment through PCR (Polymerase Chain Reaction) amplification, and inserting the target fragment into a transient expression vector to construct a gene silencing fusion expression vector; introducing the gene silencing fusion expression vector into an agrobacterium competent cell, preparing an infection solution to infect a peony plant, and injecting the infection solution into a pedicel by using an injector when a flower bud of the peony plant develops to an initial stage of a large wind chime period; when the flower buds grow to the initial stage of the round peach stage, injecting the infection liquid into pedicels and sepal bases; after each time of injection, weak light culture is carried out under a shading condition. The efficient silencing of the key gene PsuAG for regulating stamen petals is realized, and the double petals rate of the peony flower type is improved. The method does not need to depend on a stable genetic transformation system, operation is directly carried out on floral organs of a complete plant, and the gene function verification period is remarkably shortened.

The application discloses a kind of based on NPFR gene's apple snail repellent and its application, belong to the field of agricultural pest control.This application is based on RNAi-mediated NPFR gene silencing technology, design and synthesis based on NPFR gene dsNPFR, by injecting dsNPFR into apple snail, lethal effect is obvious and NPFR gene expression, weight growth rate, leaf feeding area, digestive enzyme activity etc.are significantly reduced, effectively inhibit apple snail feeding and growth, the present application provides new strategy and molecular target for RNAi biological pesticide research and development, provides new technical approach for the prevention and control of apple snail invasion hazard.

This invention relates to the field of bioengineering technology, and more particularly to a gene for identifying salt tolerance in upland cotton, GhNRX16, and its molecular marker. The CDS sequence of the upland cotton gene GhNRX16 is shown in SEQ ID NO.1 of the sequence listing. This invention uses RNA-seq technology to screen and identify this salt tolerance-related gene in the transcriptome data of upland cotton under salt stress treatment. Furthermore, the GhNRX16 sequence was obtained through gene cloning, and a transient silencing vector based on VIGS (virus-induced gene silencing) technology was successfully constructed. Experiments using VIGS-mediated gene silencing combined with salt stress treatment showed that silencing the GhNRX16 gene significantly reduced the plant's tolerance to salt stress, demonstrating that normal expression of this gene directly contributes to ensuring the plant's salt stress resistance. These results confirm the crucial role of the GhNRX16 gene in regulating cotton salt tolerance, providing important gene resources and theoretical basis for molecular breeding of salt-tolerant cotton.

The present application relates to the technical field of biological genetic engineering, and particularly relates to a kind of identification of plant salt tolerance land cotton gene GhNRX71 And molecular marker, the CDS sequence of the land cotton gene GhNRX71 As shown in SEQ ID NO.1 In the sequence table.The present application is screened and identified in the transcriptome data of land cotton under salt stress treatment by transcriptome sequencing (RNA-seq) technology.The sequence of GhNRX71 Is obtained by further gene cloning means, and the transient silencing vector based on VIGS (virus-induced gene silencing) technology is successfully constructed.The results show that the tolerance of plant to salt stress significantly reduces after silencing GhNRX71 Gene by using VIGS-mediated gene silencing combined with salt stress treatment experiment, which proves that the normal expression of the gene has a direct effect on guaranteeing the salt stress resistance of plant.The above results confirm that GhNRX71 Gene plays a key role in regulating the salt tolerance of cotton, and provides important gene resources and theoretical basis for cotton salt-tolerant molecular breeding.

The invention discloses application of a fringed iris IjFLCL1 gene to regulation and control of fringed iris growth and ecological dormancy conversion. When the fringed iris needs to be kept in an ecological dormancy state, the IjFLCL1 gene is silenced or knocked out; when the fringed iris needs to be inhibited from entering the ecological dormancy state, the IjFLCL1 gene is subjected to overexpression; the nucleotide sequence of a protein coding region of the fringed iris IjFLCL1 gene is as shown in SEQ ID No. 1. According to the invention, a homologous gene function verification method is adopted in a plant for the first time, and it is proved that silence or knockout of the IjFLCL1 gene can maintain fringed fringed iris in an ecological dormancy state. The gene has the highest expression quantity in the spring growth period and the lowest expression quantity in the dormancy induction period. Silence or knockout of the gene can delay removal of ecological dormancy of fringed iris and help plants avoid adverse effects caused by early spring climate fluctuation, so that safety and economic benefits in the cultivation process are improved.

The invention provides an MaSBP17 gene and application thereof in regulating and controlling fruit maturity, and belongs to the technical field of gene engineering. The nucleotide sequence of the MaSBP17 gene is shown as SEQ ID NO: 1 in a sequence table. According to the invention, an SBP transcription factor family in a banana genome is comprehensively and systematically analyzed, a member MaSBP17 with significant expression change in a fruit ripening process is found, and the function of the MaSBP17 is verified by using a virus-induced gene silencing technology. The invention provides an important theoretical basis and a potential molecular target for targeted regulation and control of a banana fruit ripening process, delaying of softening and aging and development of a novel postharvest preservation strategy based on key transcription factors, and has remarkable scientific value and application prospect.

The present disclosure relates to methods of creating Porcine Reproductive and Respiratory Syndrome virus (PRRSv) resistant pigs by gene silencing. In particular, the methods comprise using tools such as CRISPR / CAS9 to edit genes that either directly encode or encode molecules that are processed into non-coding RNA molecules to induce or modify their silencing activity against CD163 and / or in cells of pigs for the purpose of promoting PRRSv resistance.