13 results about "RNA silencing" patented technology

The application belongs to the field of new application of functional proteins and mutants thereof, and discloses application of sugarcane ScCBS protein and key domain deletion mutants thereof. The nucleotide sequence of the sugarcane ScCBS protein is shown as SEQ ID NO:1, and the amino acid sequence is shown as SEQ ID NO:2. The key domain deletion mutants of the ScCBS protein are ScCBS△motif1, ScCBS△motif3, ScCBS△motif4 and ScCBS△motif9, which respectively lack 28, 50, 49 and 29 amino acids. The application proves that the ScCBS protein can interact with SCSMV P1 and inhibit the RNA silencing suppressor activity by means of yeast two-hybrid, two-molecular fluorescence complementary experiment and agrobacterium transient expression system; the four mutants all lose the interaction and inhibition activity. Therefore, the ScCBS protein and the key domain deletion mutants thereof can be used for cultivating excellent SCSMV-resistant materials, and have wide application prospects in sugarcane disease-resistant molecular breeding.

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 application belongs to the technical field of genetic engineering, and particularly relates to application of cassava common mosaic virus TGBp1 in inhibiting RNA silencing. The application research finds that TGBp1 has a silencing inhibition function, can inhibit mRNA of GFP from being degraded, and thus improves the expression amount of GFP protein; it is also found that TGBp1 can inhibit a target gene MeGRXC3 from being silenced and improve the expression function thereof; in addition, it is also found that TGBp1 can improve the number of mRNA degradation pathway processing bodies (a complex formed by interaction of Dcp1 protein and Dcp2 protein) of Arabidopsis. The research result not only lays a foundation for elucidating the function of TGBp1, but also provides a new target gene for cassava disease resistance breeding.

The invention discloses a yellow dwarf disease resistance related gene MS1 and application thereof, and belongs to the technical field of biology. The invention discloses for the first time that the MS gene is a core positive regulation factor of plant broad-spectrum antiviral property through systematic research. Experiments prove that replication and diffusion of various pathogens such as BYDV, BSMV, TRV, PVX and BSCTV can be remarkably inhibited by up-regulating MS expression of plants such as wheat, barley and tobacco, and resistance does not depend on virus types. Mechanism research shows that MS silencing is achieved by enhancing PTGS-mediated RNA silencing and other basic resistance. The discovery breaks through the traditional mindset of'enhancing resistance genes', and a brand new strategy of'removing resistance inhibition factors' is created. The invention proves that MS inhibition can realize multiple virus resistance and does not damage growth and development.

The invention belongs to the fields of biotechnology, protein engineering and plant protection science, and particularly relates to an antagonistic protein mutant for cultivating a sweet potato composite virus disease (SPVD) resistant plant and application of the antagonistic protein mutant. The key pathogenic factor of the SPVD is the RNase3 protein coded by the sweet potato chlorotic stunt virus (SPCSV), and the RNase3 protein is used as an RNA silence suppressor (RSS) to destroy host immunity. The invention provides an SPCSV-RNase3 antagonistic protein mutant, which is derived from a natural antagonistic protein, namely a sweet potato trypsin inhibitor (IbSPLTI-a). According to the present invention, a compound structure model (such as construction through AlphaFold2) of wild type IbSPLTI-a and RNase3 is analyzed, and the IbSPLTI-a is modified by using a protein directed evolution strategy, particularly by using a hotspot amino acid scanning and protein surface design strategy, such that the high-activity inhibition mutant (such as IbSPLTI-a-m0805) is successfully created and screened; in-vivo and in-vitro function verification shows that the binding affinity of the mutant (such as IbSPLTI-a-m0805) and SPCSV-RNase 3 is remarkably enhanced, and the mutant shows a virus accumulation inhibition effect superior to that of a wild type IbSPLTI-a. The invention also provides a nucleic acid sequence for coding the mutant, a plant expression vector containing the sequence, and a method for culturing an anti-SPVD transgenic plant (especially sweet potato) by using the mutant. The invention provides a core gene resource and a technical path for genetic improvement of SPVD and development of a novel antiviral protein preparation.

An RNA interference-based product and method for inhibiting the expression of the double homeobox 4 (DUX4) gene on human chromosome 4q35 are disclosed. The invention comprises the silencing of the Cas13 protein of RNA, wherein Cas13 is specifically targeted to a DUX4 region of interest using a sequence-specific guide RNA (gRNA). The recombinant adeno-associated virus of the invention is constructed with a Cas13 direct repeat sequence and delivers DNA encoding an inhibitory gRNA that knocks down DUX4 expression. The method is applicable to the treatment of muscular dystrophy, including but not limited to facial-scapulohumeral muscle dystrophy (FSHD), and other disorders, including cancer, associated with elevated DUX4 expression.

RNA interference-based nucleic acids encoding double homeobox 4 (DUX4)-encoding guide RNAs (gRNA), or gRNAs that specifically hybridize to a target nucleic acid encoding DUX4 and methods for inhibiting the expression of DUX4 using the gRNAs and Cas13 are provided. The methods have application in the treatment of muscular dystrophies including, but not limited to, facioscapulohumeral muscular dystrophy (FSHD), and cancer, and other disorders associated with elevated DUX4 expression.

Provided herein are cells and compositions of cells comprising one or more nucleic acid constructs for use in assays that evaluate the delivery and potency of RNA silencing conjugates in cells.

The invention provides application of a GhMazG7 gene in regulation and control of salt tolerance of cotton, and belongs to the technical field of plant genetic engineering. The GhMazG7 gene is successfully cloned from upland cotton, the nucleotide sequence of the GhMazG7 gene is as shown in SEQ ID NO: 1, the length of an open reading frame is 465bp, the GhMazG7 gene encodes a protein containing 154 amino acids, and the amino acid sequence of the protein is as shown in SEQ ID NO: 2; according to the invention, a plant gene silencing vector aiming at the gene is constructed, then a cotton plant is transformed by using an agrobacterium tumefaciens-mediated method, the expression quantity of the endogenous GhMazG7 gene of the cotton is specifically reduced in combination with a tobacco embrittlement virus (TRV)-induced RNA silencing technology, and research finds that the expression level of the GhMazG7 gene is in positive correlation with the salt tolerance of the cotton. The method plays an important role in regulating and controlling the salt tolerance of the cotton, provides a new breeding material for further cultivating a new salt-tolerant cotton variety, and has good practicability.

This invention belongs to the field of genetic engineering technology, specifically relating to the application of Sri Lankan cassava mosaic virus BV1 in suppressing RNA silencing. This invention reveals that BV1 possesses silencing and inhibitory functions, suppressing GFP degradation and thereby increasing GFP protein expression. These findings lay the foundation for elucidating BV1 function, provide basic data for in-depth research on the infection control mechanism of SLCMV, offer new ideas for breeding disease-resistant cassava varieties, and provide new target genes for disease-resistant cassava breeding, thus playing a significant role in promoting cassava breeding.

The invention belongs to the technical field of gene engineering, and particularly relates to application of a cassava common mosaic virus TGBp1 in inhibiting RNA silencing. Research finds that TGBp1 has a silent inhibition function and can inhibit mRNA (messenger ribonucleic acid) of GFP (green fluorescent protein) from being degraded, so that the expression quantity of GFP protein is increased; the TGBp1 is also found to possibly inhibit the target gene MeGRXC3 from being silenced so as to improve the expression function of the target gene MeGRXC3; in addition, it is found that TGBp1 can increase the number of particles of an arabidopsis thaliana mRNA degradation pathway processing body (a compound formed by interaction of Dcp1 protein and Dcp2 protein), and research results not only lay a foundation for clarification of TGBp1 functions, but also provide a new target gene for cassava breeding for disease resistance.

The invention relates to newly identified nucleic acids, ribonucleic acids (RNAs) and deoxyribonucleic acids (DNAs), specifically esiRNAs / ERNAs (effective small interfering RNAs) and RNAs derived therefrom, as well as eASO (effective antisense DNA oligonucleotides), collectively referred to as eNAs (effective nucleic acids), which can be used in RNA silencing / RNAi or RNA silencinglantisense methods, as active agents against various variable plant pathogens. To identify the eNAs, an “eNA screen” was applied for the first time in standardized form to target RNAs of various plant pathogens. As a result, a new class of active agents against pathogens was identified and successfully used. The invention further relates to the construction of double-stranded ribonucleic acids, edsRNAs (effectively double-stranded RNAs), which contain nucleotide sequences of identified esiRNAs / ERNAs or related RNAs derived therefrom, and which can be successfully used in the RNA silencing / RNAi methods as active agents in plant protection against variable plant pathogens.

This disclosure relates to branched RNA compounds comprising an alkyl linker with enhanced target RNA silencing in adipose tissue.