10 results about "Silencing gene" patented technology

The invention relates to a strong promoter P7 suitable for streptomyces and application thereof, and relates to the field of genetic engineering and microbial metabolism engineering. The nucleotide sequence of the strong promoter P7 is as shown in SEQ ID No.1, and the strong promoter comprises a plasmid vector of the strong promoter; a host cell comprising the plasmid vector; the invention also discloses application of the strong promoter, the plasmid vector and the host cell in starting expression of a target gene. Compared with the prior art, the characterization of the strong promoter P7 provides an effective tool element for streptomyces strong promoter engineering and high-efficiency gene expression, and has important significance on streptomyces silent gene characterization, high-efficiency gene expression, metabolite synthesis, metabolic pathway reconstruction and the like. The strong promoter can be applied to common streptomyces type strains, and has important significance on high yield of important proteins including enzymes and important metabolites from actinomycetes.

Provided herein are compositions, systems, and methods for the creation, identification, and characterization of effector domains for activating and silencing gene expression. In particular, a high-throughput system for discovering and characterizing effector domains is provided.

This invention discloses NtBRL2 The application of genes in regulating potassium content in plants belongs to the field of plant genetic engineering technology. This invention provides a gene that can effectively regulate potassium content in plant leaves, a feature of existing technologies. NtBRL2 This invention first discovered through Real-time PCR. NtBRL2 The gene was expressed at the highest level in tobacco roots; then, a silenced gene was constructed using virus-induced gene silencing (VIGS) technology. NtBRL2 The VIGS vector of the gene was transformed into Nicotiana benthamiana, and repression was successfully achieved. NtBRL2 Silent plants, obtained by silencing gene expression, exhibit a specific phenotype with significantly lower potassium ion content compared to control plants, indicating that silencing this gene can significantly reduce potassium ion content in plants. This invention lays the foundation for elucidating the biological regulation mechanism of potassium ions and provides precise regulatory sites for the regulation of potassium ion content in plants.

A host cell comprising a first vector comprising a DNA comprising a nucleotide sequence encoding an adeno-associated virus (AAV) receptor (AAVR) or a variant thereof comprising AAVR biological activity; and a second vector comprising a DNA comprising a nucleotide sequence encoding a target gene tagged with a reporter gene is provided. A cell line comprising a plurality of the host cells comprising a first vector comprising a DNA comprising a nucleotide sequence encoding an adeno-associated virus (AAV) receptor (AAVR) or a variant thereof comprising AAVR biological activity; and a second vector comprising a DNA comprising a nucleotide sequence encoding a target gene tagged with a reporter gene is provided. A method for measuring the potency of a silencing gene therapy, the method comprising: transducing a host cell comprising a first vector comprising a DNA comprising a nucleotide sequence encoding an adeno-associated virus (AAV) receptor (AAVR) or a variant thereof comprising AAVR biological activity; and a second vector comprising a DNA comprising a nucleotide sequence encoding a target gene tagged with a reporter gene with an AAV comprising a DNA comprising a nucleotide sequence that inhibits expression of the target gene; measuring reporter gene expression in the host cell and a control; and determining a level of RNA inhibition based upon the difference in reporter gene expression in the host cell and the control is provided. In some aspects, the method is used as a potency assay to measure the potency of the AAV gene therapy product and / or measure the strength and stability of a therapeutic AAV product over time and under different storage conditions.

The application discloses an application of a leucine-rich repeat receptor-like kinase gene of Gossypium anomalum, which is composed of 3885 bases, wherein the first base from the 5' end is a transcription start site, and the 3885th base is a transcription termination site. Expression of the gene can cause a lethal phenotype of a single segment introgression line CSSL11-9 of the Gossypium anomalum. The lethal phenotype is inhibited by silencing the gene expression in the single segment introgression line CSSL11-9 of the Gossypium anomalum, and the single segment introgression line CSSL11-9 can survive normally. In addition, the gene can also endow cotton with resistance to verticillium wilt and fusarium wilt, and the resistance of the CSSL11-9 plant with the silenced gene to the cotton verticillium wilt and fusarium wilt is significantly reduced, thereby providing an important gene resource for cotton breeding against the verticillium wilt and the fusarium wilt.

This invention discloses an engineered bacterium producing tetracrine, its construction, and its application. The invention utilizes a strong promoter to drive the high expression of regulatory genes ovmZn and ovmWn in *Streptomyces neyagawa*, constructing a recombinant engineered strain Sne / pZWn; alternatively, it utilizes a strong promoter to drive the high expression of homologous genes of both, including the regulatory genes ovmZ and ovmW from *Streptomyces chromogenicus*, in *Streptomyces neyagawa*, constructing a recombinant engineered strain Sne / pZW. These engineered strains can express previously silenced gene clusters, activating tetracrine biosynthesis and laying the foundation for the development and application of tetracrine.

The invention relates to a strong promoter D8 suitable for streptomyces and application thereof, and relates to the field of genetic engineering and microbial metabolism engineering. The nucleotide sequence of the strong promoter D8 is as shown in SEQ ID No.1, and the strong promoter D8 comprises a plasmid vector of the strong promoter; the invention also discloses a host cell containing the plasmid vector, and application of the strong promoter, the plasmid vector and the host cell in starting expression of a target gene. Compared with the prior art, characterization of the promoter D8 provides a more effective tool element for streptomyces promoter engineering and high-efficiency gene expression, and the promoter D8 has important significance on streptomyces silent gene characterization, high-efficiency gene expression, metabolite synthesis, metabolic pathway reconstruction and the like. The strong promoter can be applied to common streptomyces type strains, and has important significance on high yield of important proteins including enzymes and important metabolites from actinomycetes.

The present application belongs to the field of biotechnology, and specifically relates to a method for high-throughput screening of nucleic acid drug target genes, which comprises the following steps: treating cells with nucleic acid drug candidates or negative controls, and collecting measurement values for a plurality of cell characteristic parameters related to the treatment purpose of the nucleic acid drug; calculating the fold change of the characteristic parameter measurement values of the cells in each candidate hole relative to the negative control measurement values and normalizing them; for all measurement values, calculating the squared Mahalanobis distance of the plurality of cell characteristic parameters relative to the average value using the normalized values; screening out the candidates with squared Mahalanobis distance greater than a cutoff value, and unsupervised clustering the cells according to the plurality of cell characteristic parameters to divide the candidates into at least one cluster; obtaining the corresponding silenced genes of the candidates in each cluster, and performing gene ontology enrichment analysis on the genes in the cluster to obtain the biological processes associated with each cluster, and finally obtaining nucleic acid drug target genes for different biological processes.

The present disclosure provides Crispr / cas9-based repressors for silencing gene targets in vivo and methods of use.

This invention belongs to the field of bio-agricultural technology, specifically relating to genes. StERF42 The invention relates to the application of its encoded protein in improving potato resistance to late blight. The gene discovered in this invention... StERF42 Negative regulation of potato resistance to Phytophthora in potato late blight pathogen was experimentally verified by silencing genes. StERF42 Downregulation of SA signaling pathway marker genes StPR1 The expression of this gene activates an SA-dependent immune response, enhances reactive oxygen species activity, and thus improves resistance; while overexpression of this gene... StERF42 This increases susceptibility to disease. Further research into the regulatory mechanisms revealed that genes... StERF42 StERF42 StERF42 StERF42 StERF42 StERF42 StERF42 St It possesses transcriptional activation activity and can bind to GCC-box or GGCGGTG target motifs to regulate downstream disease resistance-related genes. This invention provides a new target for potato disease resistance breeding and has broad application prospects in improving potato resistance to late blight.