35 results about "Genetic systems" patented technology

The invention relates to the technical field of plant genetic engineering, in particular to a genetic transformation method of prunus mume line plum blossom leaf calluses. The method comprises the following steps: firstly preparing a plum blossom genetic transformation receptor, then transforming agrobacterium tumefaciens by taking an RUBY gene as a reporter gene to prepare an infection bacterium solution, then carrying out callus infection and co-culture, and finally screening to obtain the resistant callus. According to the method provided by the invention, the resistant callus can be identified in an observation or PCR (Polymerase Chain Reaction) detection mode. The plum blossom callus is successfully infected, the red callus expressed by the RUBY reporter gene is obtained, the inductivity is high, and a good basis is provided for establishment and genetic improvement of a plum blossom genetic system.

The invention relates to the technical field of biotechnology and plant genetic engineering, in particular to a rice two-way single-base edited co-transcription unit gene ABE-CBE system and application of the rice two-way single-base edited co-transcription unit gene ABE-CBE system. The rice bidirectional single base edited co-transcription unit gene ABE-CBE system provided by the invention comprises a co-transcription unit gene ABE-CBE, and the co-transcription unit gene ABE-CBE comprises an encoding gene ABE8e of adenine deaminase and an encoding gene Sdd7 of cytosine deaminase, and the coding gene ABE8e and the coding gene Sdd7 in the co-transcription gene ABE-CBE are constructed in the same expression vector. Through the designed ABE-CBE system, the two editors can be mediated at the same site to play a role at the same time, generation of unexpected base substitution can be further reduced, the editing efficiency of the base editors is improved, and wide application value is achieved.

Cas-cleavable reporter systems, CRISPR-Cas systems thereof, and methods of use thereof in CRISPR-Cas based diagnostics. Cas-cleavable reporter systems may generate a luminescent, fluorescent, or other detectable signal upon Cas-collateral cleavage of one or more reporter system components. CRISPR-Cas systems may comprise a Cas protein having collateral cleavage activity, guide molecules, and the Cas-cleavable reporter system. Cas-cleavable reporter systems may be a split luciferase reporter system, at least one element of which is bound to beads. For multiplexing, CRISPR-Cas systems may comprise a Cas-cleavable quenched reporter system, optionally a Cas-cleavable quenched fluorescent reporter system, a Cas protein having collateral cleavage activity, and target-specific guide molecules attached to color-coded beads. CRISPR-Cas systems may further comprise amplification reagents. Methods may apply said CRISPR-Cas systems to flow cell, well-plate, or other devices, and may measure detectable signals by microscopic, plate reader, or other methods.

The invention belongs to the technical field of bioinformatics, and particularly relates to a method for screening cross-species HGT. The method comprises the steps of constructing a background database, inferring a homologous group, constructing a gene phylogenetic tree, inferring HGT based on the gene tree, inferring verification of candidate HGTs based on the gene tree, distributing the HGTs to a timeline and the like. According to the method, high-throughput and automatic screening of HGT events in a large number of gene datasets is realized by integrating a plurality of steps of phylogenetic analysis, sequence alignment, gene function annotation, statistical verification and the like. The method disclosed by the invention is particularly suitable for the HGT research of an ultra-long time length and an ultra-long evolution distance crossing a biological boundary, and an important scientific tool can be provided for understanding gene flow and function enhancement in a biological evolution process.

The invention discloses a method for creating a high-frequency introgression Chinese cabbage-cabbage introgression line, and relates to the technical field of introgression breeding. According to the method, tetraploid Chinese cabbage and diploid cabbage are subjected to interspecific hybridization to obtain an allotriploid hybrid (AAC), then the AAC is subjected to hybridization and selfing with diploid Chinese cabbage, and a Chinese cabbage-cabbage introgression line with a high-frequency introgression exogenous species genome sequence is created in combination with free microspore culture. According to the invention, the heterotriploid hybrid mediated genetic system is utilized to realize interspecific high introgression, and a new method is provided for breaking the bottleneck of low exogenous species introgression frequency.

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 invention discloses a novel molecular chaperone-mediated autophagy (CMA) activating agent, relates to the technical field of biological cells, and is characterized in that the novel molecular chaperone-mediated autophagy activating agent is a novel anti-tumor drug chidamide, and the novel molecular chaperone-mediated autophagy activating agent is a novel anti-tumor drug chidamide. The chidamide can obviously up-regulate the mRNA and protein level of the key protein LAMP2A in the CMA process. According to the present invention, by constructing a human THP1-LAMP2luciferase reporter gene system, the compound capable of activating the LAMP2 promoter is screened from the bulk drug; through cell level primary screening, secondary screening and animal level verification, an effective CMA activator chidamide is finally determined. A good tool is provided for basic research of CMA, and novel drugs are further researched and developed by taking CMA as a target spot.

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.

The utility model provides a wireless photogenetic system of a charging version, which solves the problems that the activity range of an animal is limited and the experimental design is influenced in the existing photogenetic experiment through an animal head connection mode, and the main scheme is that the wireless photogenetic system comprises a control host and a plurality of implantable photoelectric probes, the plurality of implantable photoelectric probes are independently packaged and can be independently controlled through the control host, the plurality of implantable photoelectric probes can be charged through the charging box at the same time, each implantable photoelectric probe comprises an RF chip, an antenna, a battery, a flexible probe substrate and a miniature LED, and the front and back surfaces of the flexible probe substrate are provided with a plurality of groups of joints. The micro LEDs are electrically connected with the connectors in a one-to-one correspondence mode, the battery is electrically connected with the RF chip and the micro LEDs, the RF chip is electrically connected with the micro LEDs and transmits external signals through the antenna, and the control host adjusts output of the micro LEDs through the driving circuit.

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 invention discloses a synsepalum dulcificum protein coding gene endogenous promoter and application thereof. The endogenous promoter of the synsepalum dulcificum protein coding gene is any one of proSdMIR.1, proSdMIR.1.2, proSdMIR.1.3 and proSdMIR.1.4, the DNA (deoxyribonucleic acid) nucleotide sequence of the proSdMIR.1 is as shown in SEQ ID NO.3, the DNA nucleotide sequence of the proSdMIR.1.2 is as shown in SEQ ID NO.4, the DNA nucleotide sequence of the proSdMIR.1.3 is as shown in SEQ ID NO.5, and the DNA nucleotide sequence of the proSdMIR.1.4 is as shown in SEQ ID NO.6. The synsepalum dulcificum protein coding gene disclosed by the invention has the advantages that the synsepalum dulcificum protein coding gene can be used for preparing the synsepalum dulcificum protein The miraculin gene promoter provided by the invention has important significance in optimizing a miracle fruit genetic system and creating miracle fruit germplasm resources with excellent characters.

The present invention belongs to the field of bioinformatics technology and specifically relates to a method for screening cross-species HGT. The method includes the steps of constructing a background database, inferring homology groups, constructing a gene phylogenetic tree, inferring HGT based on the gene tree, verifying candidate HGT inferred based on the gene tree, and assigning HGT to a timeline. By integrating a multi-step process such as phylogenetic analysis, sequence alignment, gene function annotation, and statistical verification, the method achieves high-throughput, automated screening of HGT events in large genetic datasets. The method of the present invention is particularly suitable for studying HGT across extremely long time periods and extremely long evolutionary distances across the biological kingdom, and can provide an important scientific tool for understanding gene flow and functional enhancement in the course of biological evolution.

The invention discloses a subtropical estuary benthic fish mullet chubil AHR1b gene recombinant expression plasmid and application thereof, and belongs to the technical field of environmental toxicology. The AHR1b gene recombinant expression plasmid is prepared by introducing an AHR1b gene into an expression vector. The invention also discloses a luciferase reporter gene system, which comprises an AHR1b gene recombinant expression plasmid, an ARNT1 gene recombinant expression plasmid and a firefly luciferase reporter plasmid containing a mouse CYP1A1 gene promoter. According to the invention, a luciferase reporter gene system is utilized to carry out comprehensive characterization on TCDD toxicity and evaluate the sensitivity and the specificity of a mugilogobius chuchuyi AHR1b and ARNT1 subtype dependency reporter gene system, so that a theoretical basis is provided for supporting high-throughput and high-sensitivity environment monitoring, and meanwhile, a species-specific early warning technology is provided for offshore and estuary DLCs pollution.

The invention discloses an enhancer for regulating and controlling chicken PGC-1alpha gene expression and application of the enhancer, and belongs to the field of bioengineering. The nucleotide sequence of the enhancer is shown as SEQ ID NO: 1. The invention further discloses a biological material containing the enhancer and a preparation method of a cell strain with the enhancer knocked out. A dual-luciferase reporter gene system proves that the enhancer has remarkable activity in DF1 and ICP2 cells, the expression of the PGC-1alpha gene in chicken preadipocytes can be reduced by knocking out the enhancer, and cell differentiation and lipid droplet generation are remarkably promoted. The enhancer can be used for preparing a biological preparation for regulating and controlling the expression level of the PGC-1alpha gene and regulating and controlling chicken preadipocyte differentiation, can also be applied to genetic improvement of chicken fat traits and can be used for preparing a model for researching related gene functions and cell differentiation mechanisms. The invention provides a new target and technical support for chicken fat character improvement.

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 utility model provides a radio frequency-based wireless optical genetic system, which solves the problems that in the existing optical genetic stimulation system, most flexible implants depend on an external battery module to supply power, the service time is limited and the like, and the main scheme is that the radio frequency-based wireless optical genetic system comprises a radio frequency emission source, a radio frequency switch, an impedance coupling adjusting circuit, the flexible implants, a touch screen and a case, the radio frequency emission source can generate a radio frequency signal with a specific frequency; the radio frequency switch is electrically connected with the radio frequency emission source so as to control the on-off of the radio frequency emission source; the impedance coupling adjusting circuit is used for adjusting impedance matching with the radio frequency emission source according to different coil sizes; the whole flexible implant is made of a flexible circuit board; the touch screen is used for being matched with control software to set optical genetic parameters; and the case integrates the radio frequency emission source and the radio frequency switch into a whole machine.

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 present invention provides a method for establishing an Agrobacterium-mediated evergreen blueberry transgenic system, comprising: adding a rhizogenes containing a functional gene to a liquid culture medium to prepare an infection solution; a vacuum infiltration treatment method is: placing an explant in the infection solution for vacuum infiltration treatment, and then placing it on a co-culture medium for co-culture; using the tender shoots of evergreen blueberry seedlings grown under sterile conditions as explants, placing the explants in the infection solution for vacuum infiltration treatment, and then placing them on a co-culture medium for co-culture; washing the explants after co-culture, and then placing them on a regeneration medium for regeneration culture; after 15d to 20d of regeneration culture, the explants gradually grow transgenic callus, and buds appear, and finally the evergreen blueberry transgenic system is obtained. The present invention realizes the transgenic of evergreen blueberry callus for the first time, and obtains transgenic buds, with a conversion rate of 3.27%, and it only takes 2-3 months to obtain transgenic buds.

The application discloses a subtropical estuary benthic fish Oxyeleotris marmoratus AHR1b gene recombination expression plasmid and application thereof, and belongs to the technical field of environmental toxicology. The AHR1b gene recombination expression plasmid is prepared by introducing the AHR1b gene into an expression vector. A luciferase reporter gene system is also disclosed, which comprises the AHR1b gene recombination expression plasmid, an ARNT1 gene recombination expression plasmid and a firefly luciferase reporter plasmid containing a mouse CYP1A1 gene promoter. The luciferase reporter gene system is used to comprehensively characterize TCDD toxicity, to evaluate the sensitivity and specificity of the Oxyeleotris marmoratus AHR1b and ARNT1 subtype-dependent reporter gene system, to provide a theoretical basis for supporting high-throughput and high-sensitivity environmental monitoring, and to provide a species-specific early warning technology for offshore and estuary DLCs pollution.

The invention discloses an AHR2a and ARNT1 gene recombinant expression plasmid of subtropical estuary benthic fish mullet chubil and application of the recombinant expression plasmid, and belongs to the field of environmental toxicology. According to the recombinant expression plasmids, AHR2a and ARNT1 genes are respectively connected with an expression vector to construct a recombinant expression plasmid I and a recombinant expression plasmid II. The invention also constructs a luciferase reporter gene system containing the two recombinant expression plasmids and a firefly luciferase reporter plasmid containing a mouse CYP1A1 gene promoter. By utilizing the luciferase reporter gene system, the toxicity of dioxin and dioxin-like chemical substances and complex environmental wastewater can be comprehensively represented, the sensitivity and the specificity of the mugilogobius chuenensis AHR2a and ARNT1 subtype dependency reporter gene system are evaluated, and a theoretical basis is provided for supporting high-throughput and high-sensitivity environmental monitoring; and a species-specific early warning technology is provided for offshore and estuary DLCs pollution.

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 invention provides a construction method of a fluorescent tracing mouse and application of the fluorescent tracing mouse in sorting Mcpt4 positive and negative mast cells. According to the invention, a Cre gene is knocked into an Mcpt4 gene sequence of a C57BL / 6 mouse by using a CRISPR / Cas9 technology, and then a fluorescence tracing mouse model of Mcpt4 positive mast cells is obtained through animal hybridization and a Cre-loxP recombination technology. The fluorescent tracing mouse model has a red fluorescent protein tdTomato and green fluorescent protein ZsGreen double-fluorescence reporter gene system, Mcpt4 positive mast cells show red fluorescence, and other Mcpt4 negative cells show green fluorescence. By utilizing the fluorescence tracing mouse model, the Mcpt4 positive and negative mast cells can be quickly, accurately and effectively sorted, and a reliable technical basis is provided for exploring the action and mechanism of the Mcpt4 positive and negative mast cells in immunoreaction.

The invention discloses a method for knocking out a bovine FBXO40 gene and application of the method, and belongs to the technical field of animal bioengineering. A CRISPR (clustered regularly interspaced short palindromic repeats) / Cas12i system is used for knocking out the bovine FBXO40 gene, the CRISPR / Cas12i system targets a fifth exon of the bovine FBXO40 gene, and a nucleotide sequence of the fifth exon of the bovine FBXO40 gene is as shown in SEQ ID No. 1. The invention provides a method for realizing accurate knockout of the cattle FBXO40 gene by utilizing a CRISPR / Cas12i gene editing technology, constructs a gene knockout monoclonal cell line and embryo capable of stably inheriting, can provide experimental reference for research of a cattle skeletal muscle development molecular mechanism, and can also be applied to cultivation of a new variety of cattle with high meat yield.

The invention belongs to the technical field of biology, and particularly relates to a system for improving circular RNA expression in mammalian cells. The construction method of the system plasmid comprises the step of constructing the optimized Tornado system plasmid by sequentially connecting a luciferase gene, a Tornado system cyclization element, a circRNA translation expression cassette and a target gene. According to the present invention, by prolonging the distance between the CMV promoter and the IRES, and increasing the 5 'LTR (including the upstream RSV promoter) and the WPRE, the expression cassette capable of significantly improving the protein expression level of the IRES-containing exogenous circRNA is constructed, and the RNA cyclization system can be used for expressing the exogenous functional target protein so as to be used for the research and development of the therapeutic drug or the prophylactic vaccine.