16 results about "Genetic systems" patented technology

This invention provides a method for constructing a fluorescently traced mouse model and its application in the sorting of Mcpt4-positive and negative mast cells. First, the Cre gene is knocked into the Mcpt4 gene sequence of C57BL / 6 mice using CRISPR / Cas9 technology. Then, through animal hybridization and Cre-loxP recombination technology, a fluorescently traced mouse model of Mcpt4-positive mast cells is obtained. The fluorescently traced mouse model possesses a dual-fluorescent reporter gene system consisting of the red fluorescent protein tdTomato and the green fluorescent protein ZsGreen. Mcpt4-positive mast cells exhibit red fluorescence, while other Mcpt4-negative cells exhibit green fluorescence. Using this fluorescently traced mouse model, Mcpt4-positive and negative mast cells can be rapidly, accurately, and effectively sorted, providing a reliable technical basis for exploring the roles and mechanisms of Mcpt4-positive and negative mast cells in immune responses.

The present invention provides an in-vitro method of reducing the efficiency of transducing malignant cells of the blood system of a subject that are not derived from T cells with lentiviral vector particles without reducing the efficiency of transducing T cells in a sample comprising T cells and said malignant cells. A combination of compositions comprising a first composition and a second composition is also disclosed, wherein said first composition comprises i) transduced T cells of a subject, wherein said transduced T cells express a CAR comprising an antigen binding domain, wherein the antigen binding domain of said CAR binds specifically to a tag of a tagged polypeptide, and ii) non-transduced malignant cells of the blood system of said subject, and wherein said second composition comprises said tagged polypeptide, wherein said tagged polypeptide binds specifically to an antigen expressed on the surface of said malignant cells. Alternatively, the transduced T cells of said first composition may comprise a nucleic acid encoding a CAR and an inducible gene expression system, and said second composition may comprise an induction agent inducing said gene system.

The invention discloses a screening and action mechanism research method for anti-pneumonia active ingredients of Xiaobai powder, and relates to the technical field of biological medicines, and the technical key points are as follows: three acute pneumonia animal models simulating the inducement characteristics of'infantile pneumonia 'are constructed, so that XBS anti-pneumonia efficacy evaluation is more representative; meanwhile, preliminary screening is carried out by applying a spectrum-activity correlation modeling technology, then pharmacodynamic active ingredients of XBS for treating pneumonia are screened by virtue of an anti-inflammatory drug luciferase reporter gene system of an NF-kappa B signal channel, so that the subsequent analysis work is more concentrated, and a Poly-PK / PM-PD comprehensive strategy is adopted, so that the accuracy of screening is improved. The pharmacodynamic action mechanism of XBS for treating pneumonia is explained from dynamic change correlation of four groups of quantitative data, a new thought is provided for treating infantile lung heat cough and asthma, and meanwhile, the characteristics of progressive and multi-path effects of a compound traditional Chinese medicine are also shown.

This invention discloses an enhancer that regulates chicken PGC-1α gene expression and its applications, belonging to the field of bioengineering. The nucleotide sequence of the enhancer is shown in SEQ ID NO: 1. This invention further discloses biomaterials containing this enhancer and a method for preparing cell lines with the enhancer knocked out. A dual-luciferase reporter gene system confirmed its significant activity in DF1 and ICP2 cells. Knocking out this enhancer downregulates PGC-1α gene expression in chicken preadipocytes, significantly promoting cell differentiation and lipid droplet formation. This enhancer can be used to prepare biological agents that regulate PGC-1α gene expression levels and chicken preadipocyte differentiation. It can also be applied to the genetic improvement of chicken fat traits and to the preparation of models for studying related gene function and cell differentiation mechanisms. This invention provides new targets and technical support for improving chicken fat traits.

The invention belongs to the technical field of medical chemistry, and discloses an aza aromatic amine protein degradation targeting chimera (PROTAC) compound with the activity of degrading nuclear receptor co-inhibitor 1 (NCoR1) in a targeting manner, and a preparation method and application of the aza aromatic amine protein degradation targeting chimera (PROTAC) compound. The aza aromatic amine compound is prepared through acylation and N-alkylation reaction, and the structure of the compound is shown as a formula I to a formula IV. A Liver X Receptor Response Element-Luciferase reporter gene system (LXRE-Luc luciferase reporter gene system) determination experiment shows that when the compound is 10 [mu] M, the compound has an obvious liver receptor (LXR) regulation effect, and the compound has an obvious liver receptor regulation effect when the compound is 10 [mu] M; western blot experiment research shows that the compound has a remarkable degradation effect on NCoR1, and the compound XH-10 with the highest activity can generate a remarkable NCoR1 degradation effect when the concentration is 5 mu M. The compound is PROTAC with NCoR1 targeted degradation activity, and has the advantages of high activity and simple preparation process. The compound has an important value in the aspect of glycometabolism and ester metabolism regulation medicine development.

The invention relates to the technical field of bioengineering, and discloses a construction method and application of recombinant Escherichia coli engineering bacteria with high yield of ergothioneine, and the method comprises the following steps: by taking Escherichia coli as a chassis, integrating a mycobacterium smegmatis egtABCDE gene cluster at an ldhA site; a repressor protein gene metJ and a cysteine lyase gene yhaM are knocked out, a methionine adenosine transferase gene metK and a serine acetyltransferase gene cysE are overexpressed by using a strong promoter, and mutant metA, thrA, serA and hisG genes for relieving feedback inhibition are introduced, so that metabolic inhibition of methionine, cysteine and histidine pathways is systematically relieved; the invention also establishes a matched fermentation process. According to the invention, endogenous precursor supply and exogenous synthesis pathways are synergistically enhanced, the precursor supply bottleneck is solved, the yield of ergothioneine is increased, and the method is suitable for industrial production.

The application discloses a miraculin gene endogenous promoter and application thereof. The miraculin gene endogenous promoter is any one of proSdMIR.1, proSdMIR.1.2, proSdMIR.1.3 and proSdMIR.1.4, the DNA nucleotide sequence of proSdMIR.1 is shown as SEQ ID NO.3, the DNA nucleotide sequence of proSdMIR.1.2 is shown as SEQ ID NO.4, the DNA nucleotide sequence of proSdMIR.1.3 is shown as SEQ ID NO.5, and the DNA nucleotide sequence of proSdMIR.1.4 is shown as SEQ ID NO.6. The miraculin gene promoter provided by the application is of great significance for optimizing a genetic system of miraculin and creating a miraculin germplasm resource with excellent traits.

The present invention provides high-content and high-resolution in vivo screens for mapping genes that are linked to changes in cytoarchitecture or spatial transcriptomics of brain cells (e.g., neurons). The multiplex gene mapping platform utilizes genetic perturbations that are delivered to a CRISPR-expressing transgenic system via viral vectors (e.g., lentiviral vectors or AAV vectors). Phenotypic changes in the cytoarchitecture or spatial transcriptomics resulting from genetic perturbations are examined in individual brain cells, which can then be correlated with specific target genes that are modulated by the genetic perturbations.

The disclosure belongs to the technical field of animal bioengineering. Disclosed is a method for knocking out a bovine FBXO40 gene and an application thereof. A CRISPR / Cas12i system is used to knock out the bovine FBXO40 gene. The CRISPR / Cas12i system targets exon 5 of the bovine FBXO40 gene, and a nucleotide sequence of the exon 5 of the bovine FBXO40 gene is shown in SEQ ID No. 1. In the disclosure, a method for precise knockout of the bovine FBXO40 gene using a CRISPR / Cas12i gene editing technology is provided, and stably inheritable monoclonal cell lines and embryos with gene knockouts are successfully constructed. The method can provide experimental references for studying a molecular mechanism for bovine skeletal muscle development, and can also be applied in the breeding of new cattle breeds with high-meat-yielding.

The present disclosure relates to a chemogenetic system and method treating endometriosis using a gene therapy vector to deliver a therapeutic payload to specific cells (nociceptive neurons). Overexpression of the payload increases the sensitivity of the cells to an activator, thereby enabling lower doses of activator drugs to be administered. The disclosure also related to vectors to deliver a nucleotide sequence to the target cells to enable overexpression thereof.

The application discloses a luciferase reporter gene system and application thereof in screening of active ingredients of traditional Chinese medicines, and relates to the technical field of biological medicine and molecular biology. The application claims a luciferase reporter gene system, which can be specifically used for screening and finding YAP-TEAD complex transcription activity inhibitors, and especially can be used for screening target compounds capable of significantly inhibiting YAP-TEAD complex transcription activity from traditional Chinese medicines such as Alisma orientalis (Lour.) Juzep and traditional Chinese medicine compound prescriptions. The compounds obtained through the screening can be used for clarifying the material basis of the pharmacological effects of the traditional Chinese medicines such as Alisma orientalis (Lour.) Juzep and traditional Chinese medicine compound prescriptions, and determining the molecular mechanism of the pharmacological effects, and can be used as candidate compounds for development of new drugs, and provide important theoretical basis and material support for subsequent research and development of drugs for resisting related diseases.

The present disclosure relates to a chemogenetic system and method of treating Bladder Musculature dysfunction (BMD) comprising using a gene therapy vector that selectively transduces lower urinary tr

The invention provides a construction method of a monoclonal cell strain for knocking out a Tan sheep FABP4 gene, which comprises the following steps: knocking out the Tan sheep FABP4 gene by using a CRISPR / Cas9 system, targeting a first exon of the Tan sheep FABP4 gene by using the CRISPR / Cas9 system, and showing a nucleotide sequence of the first exon of the Tan sheep FABP4 gene as SEQ ID NO.1; sgRNA in the CRISPR / Cas9 system targets the nucleotides at the 2-8 sites at the 5'end in the sequence SEQ ID NO.1, and the nucleotide sequence of the sgRNA is as shown in SEQ ID NO.2; the method comprises the following steps: constructing a Lentiti-CRISPR V2-Puro-OaFABP4-sgRNA recombinant plasmid, and carrying out lentivirus packaging, drug screening and monoclonal screening, so as to obtain the monoclonal cell strain of which the Tan sheep FABP4 gene is knocked out. The invention also provides application of the monoclonal cell strain, and the monoclonal cell strain is used for constructing a cell model of a Tan sheep precursor fat cell differentiation regulation mechanism. According to the invention, the Tan sheep FABP4 gene is knocked out by using a CRISPR / Cas9 system, so that the research on the function and regulation mechanism of the FABP4 gene in Tan sheep precursor fat cell differentiation is facilitated, and a new molecular target is provided for controlling excessive fat accumulation.

The invention belongs to the technical field of medical chemistry, and discloses a secondary amine PROTAC compound with the activity of degrading a nuclear receptor co-inhibitor 1 (NCoR1) in a targeted manner as well as a preparation method and application of the secondary amine PROTAC compound. The secondary amine compound is prepared through acylation and N-alkylation reaction, and the structure of the compound is shown as a formula I to a formula IV. A Liver X Receptor Response Element-Luciferase reporter gene system (LXRE-Luc luciferase reporter gene system) determination experiment shows that when the compound is 10 [mu] M, the compound has an obvious liver receptor (LXR) regulation effect, and the compound has an obvious liver receptor regulation effect when the compound is 10 [mu] M; western blot experiment research shows that the compound has a remarkable degradation effect on NCoR1, and the compound ZD-5 with the highest activity can generate a remarkable NCoR1 degradation effect when the concentration is 5 mu M. The compound is a protein degradation targeting chimera (PROTAC), has the advantages of being high in activity and easy and convenient to prepare, and has great value in the aspect of glycometabolism and ester metabolism regulation medicine development.

The invention belongs to the technical field of molecular number inheritance and molecular breeding, and particularly relates to a multi-character index model construction method based on crop cross breeding optimization combination design. The method comprises the following steps: carrying out genetic system analysis by utilizing whole genome sequencing data to obtain gene loci for controlling quantitative characters and effects thereof, further establishing a gene-allele matrix of multiple characters of a breeding population, then carrying out genome simulation on homozygous offspring population of all potential parent combinations in the breeding population, and finally carrying out genetic system analysis on the homozygous offspring population. And estimating the genotype value of the offspring individual according to the gene-allele matrix. And finally, constructing a breeding value selection index model of multiple traits, establishing a breeding potential evaluation system of the parent combination by integrating multi-trait information, and carrying out parent combination optimization design, so as to screen out an excellent hybrid combination and improve the plant breeding efficiency.