82 results about "Eukaryotic organism" patented technology

The invention belongs to the field of gene engineering, and relates to a DNA binding protein based on TraC effect protein, a fusion protein with a cytosine / adenine base editing function, an editing system and nucleic acid molecules or constructs for encoding the DNA binding protein and the fusion protein. The invention also relates to complexes and kits for gene editing comprising the fusion proteins of the invention or the nucleic acid molecules encoding them. The base editing tool disclosed by the invention can realize efficient, accurate and safe base editing in cells of various eukaryotes such as animals and plants. The unique characteristics of the base editing tool enrich the types of existing base editing tools, and the base editing tool has wide application prospects.

The invention belongs to the technical field of gene engineering, and particularly relates to a biotin ligase mutant as well as a coding gene, an expression vector, a recombinant cell and application thereof. The amino acid sequence of the biotin ligase mutant is as shown in SEQ ID NO. 2. The biotin ligase (Bar A) mutant provided by the invention can be efficiently expressed in eukaryotes, the expression level of the biotin ligase (Bar A) mutant is obviously improved in mammalian cells compared with a wild type, and the biotin ligase (Bar A) mutant has good catalytic activity after expression, can effectively catalyze biotinylation of protein containing a biotin receptor sequence, and has good application prospects. The method has a wide application prospect in the fields of biotinylated protein preparation, and protein labeling, positioning, immunodetection, affinity purification, function research and the like based on a biotin-avidin system (BAS).

The present invention relates to a modified stem cell and an ex vivo method for preparing defined mature cells from genetic programming of said modified stem cell which can be carried out in absence of certain growth factors, as well as a kit allowing to transform stem cells, and the different uses of the mature cells, for example for in vitro drug screening and toxicology or as cultivated meat. The genetic programming involves a molecular switch between two mutually exclusive programs of stem cell expansion and differentiation. The invention may be used for stem cells or other progenitors cells of any type, from any eukaryotic organism, but finds particular application in pluripotent stem cells from humans and livestock animals.

The present invention generally relates to systems (100) and methods for generating, managing, and displaying non-fungible digital representations (102) of living, biological, and / or eukaryotic organisms (104). More specifically, the systems (100) and methods for generating, managing, and displaying non-fungible digital representations (102) include genetic material (106), such as single nucleotide polymorphisms. The systems (100) and methods can record genetic data from the genetic material (106) on a blockchain (114) and can generate and display the non-fungible digital representation (102) of the living, biological, and / or eukaryotic organisms (104) that are based on the genetic data on the blockchain (114).

The invention relates to the technical field of bioinformatics and molecular cytogenetics, in particular to a method for counting centromeres of chromosomes based on a Nuf2 antibody and application. The method comprises the following steps: performing chromosome centromere counting on a Nuf2 antibody and a centromere specific protein Nuf2 through an immunofluorescence reaction, so that the number of chromosome centromeres can be directly observed, and the number of chromosomes can be efficiently and quickly determined. Besides, due to the fact that the specific protein Nuf2 of the centromere is highly conservative in eukaryotes, the invention further provides application of the method to identification of the plant ploidy, the method is easy and convenient to operate and low in cost, the ploidy identification efficiency is remarkably improved, and a universal technical platform is provided for genetic breeding.

The present invention provides a versatile platform system for regulating gene expression in cells of eukaryotic organisms, including plants, algae and fungi, and mammals. A platform system of the invention includes an expression control element and expression system comprising the same. Expression control elements according to the invention can be inserted into a variety of cell types having the capability of alternative splicing and can be regulated by a polyamine or polyamine analogues. Host cells comprising the expression control elements of the invention are typically grown in a cell or tissue culture, or form part of a living plant, algae, or fungi.

The present invention relates to Split-Intron Final Self-Inactivating (SIN) retroviral vectors which comprises a viral major splice donor (mSD) comprising mutations, a mouse mammary tumor vector long terminal repeat (MMTV-LTR) or a Type B leukemogenic virus long terminal repeat (TBLV-LTR), a eukaryotic splice acceptor (eSA), and a eukaryotic splice donor (eSD). Furthermore, the invention relation to the use and methods of uses of such vectors in gene therapy.

The invention provides a SpuFz1 variant derived from eukaryotes and application of the SpuFz1 variant in gene editing, particularly provides the SpuFz1 variant and modified omega RNA, and finds that the modified omega RNA and SpuFz1 mutant combination can remarkably improve the editing efficiency and greatly reduce the off-target rate for the first time.

The invention discloses base-modified mRNA (messenger Ribonucleic Acid) and application thereof in a cell-free protein synthesis system, and belongs to the technical field of nucleic acid modification. Natural nucleoside triphosphate is completely replaced by different types of basic group modified nucleoside triphosphate, mRNA with different chemical modifications is obtained through an in-vitro transcription method and purification, and the mRNA is put into an in-vitro cell-free protein synthesis system for reaction. A reaction element recombination system-PURE system based on prokaryote Escherichia coli purified protein and an eukaryote CFPS system taking rabbit reticulated red lysate as a material are mainly selected to detect the corresponding translation efficiency of mRNA modified by different bases in two cell-free protein synthesis systems respectively. Different regulatory activities may be shown in different cell-free protein synthesis systems when different modified nucleotides are doped into mRNA, base modified triphosphoric acid capable of improving the translation efficiency of an in-vitro protein synthesis system is screened out, and the stability of mRNA in the cell-free protein synthesis system and the corresponding protein expression level are improved.

The Quadruplet Expanded DNA (QED) eukaryote genetic code comprising twenty nondegenerate QED codons encode proteins (the protein-encoding codons), and thirty-five nondegenerate QED codons (the noncoding codons) being highly correlated with cis-regulatory elements control and regulate transcription, alternate splicing, and polymerization in eukaryotic protein synthesis using canonical amino acids. The QED eukaryote genetic code is an advancement to gene therapeutics that allows for the correction of dysfunctional proteins. Additionally, the QED eukaryote genetic code is further applicable for changing paradigms relating to identifying cures for monogenic rare, multigene cancer, and neurodegenerative diseases.

An engineered tyrosyl-tRNA synthetase / tRNA pair that co-translationally incorporates O-sulfotyrosine in response to UAG codons in E. coli and mammalian cells is described herein. This platform enables recombinant expression of eukaryotic proteins homogeneously sulfated at chosen sites.

Novel combinations of cysteine-rich insecticidal proteins (CRIPs) and insecticides (IAs), such as chemicals, molecules, nucleotides, polynucleotides, peptides, polypeptides, proteins, toxins, toxic substances, poisons, insecticides, pesticides, organic compounds, inorganic compounds, prokaryotes or eukaryotes (and substances produced from such prokaryotes or eukaryotes), are described and claimed for the control and / or eradication of pests. [Solution] Novel insecticide combinations, compositions, and methods for using them are provided. The present invention relates to a combination of a cysteine-rich insecticide peptide (CRIP) and an insecticide (IA). The present invention also describes the use of the combination and composition for controlling insects. This specification describes the gene encoding CRIP, compositions and combinations containing CRIP and IA, and methods useful for controlling pests using them.

A method for improving the replication of covalent closed circular plasmids is provided. [Solution] The method comprises the step of preparing a covalent closed circular plasmid having a Pol I-dependent origin of replication and an insert containing a structured DNA sequence selected from the group consisting of a reverse repeat sequence, a directional repeat sequence, a homopolymer repeat sequence, a eukaryotic origin of replication, or a eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also comprises the step of modifying the covalent closed circular recombinant molecule so that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, thereby improving the replication of the covalent closed circular plasmid with the resulting Pol III-dependent origin of replication. A covalent closed circular recombinant DNA molecule without antibiotic markers is also provided.

Disclosed are split complementary base editing systems based on bimolecular deaminases and uses thereof. A split complementary base editing system mainly includes base editing fusion proteins A and B that are from splitting at a deaminase domain embedded inside a Cas9 nickase (nCas9), and a guide RNA (gRNA). The present disclosures of split complementary cytosine base editing systems can greatly reduce Cas9-dependent and Cas9-independent off-target effects in the genome while maintaining robust on-target cytosine base editing. The present disclosures also provide the split complementary adenine base editing systems. The split complementary cytosine (adenine) base editing systems are widely applicable to various eukaryotic organisms, and can be used in crop genetic breeding, animal breed improvement, and even clinical treatment of human genetic diseases.

The present invention. relates to a disinfectant composition against parasitic organisms. The invention also relates to a method for disinfecting at least one area to prevent parasitosis. The invention also relates to the use of at least one compound selected from fatty acids and fatty alcohols and which have a chain length of to carbon atoms in a disinfectant composition for improving penetration of active compounds contained in said composition into the durable forms, in particular eggs or oocysts, of parasitic organisms.

The present invention relates to an oligonucleotide comprising: a) a nucleotide sequence of a broom-kojin-ricin loop (SRL) of a eukaryotic / mammalian 60S ribosomal subunit wherein the SRL nucleotide sequence comprises at least one adenine; and b) at least one cut-dependent marker; wherein the oligonucleotide is a single strand. Preferably, the oligonucleotides form at least one loop structure or stem-loop structure. In another aspect, the invention relates to a method of detecting active shiga toxin in a sample, the method comprising the steps of: a) providing at least one oligonucleotide according to the invention; b) providing a sample of shiga toxin to be detected; c) incubating the sample with at least one single-stranded oligonucleotide; and d) detecting a labeled signal, wherein the signal indicates the presence of Shiga toxin in the sample. The invention additionally comprises a kit comprising at least one oligonucleotide according to the invention.

Provided are: a nucleic acid cleavage enzyme which comprises a nuclease domain having a specific amino acid sequence and a nucleic acid binding domain; a use of the nucleic acid cleavage enzyme; a method for producing a variant of a eukaryotic cell, the method comprising deleting a centromere region of a genome on a chromosome in the eukaryotic cell; a use of the method; a vector or a DNA fragment for use in engineering of a gene of a eukaryotic organism, the vector or the DNA fragment including a telomeric repeat sequence; and a use of the vector or the DNA fragment.

The present invention relates to a disinfectant composition against parasitic organisms. The invention also relates to a method for disinfecting at least one area to prevent parasitosis. The invention also relates to the use of at least one compound selected from fatty acids and fatty alcohols and which have a chain length of 10 to 26 carbon atoms in a disinfectant composition for improving penetration of active compounds contained in said composition into the durable forms, in particular eggs or oocysts, of parasitic organisms.

The present application provides a kind of eukaryotic pan-transcriptome annotation method, comprising the following steps: the transcriptome data of more than 3 different sources of eukaryotic species sample is compared to reference genome, obtains corresponding gene structure annotation information, integrates, obtains the transcriptome annotation result of reference genome dependence;Sample transcriptome data is spliced and assembled, and corresponding mature transcript sequence information is obtained, and the mature transcript sequence information is compared with reference genome, and corresponding gene structure information is obtained, to obtain the transcriptome annotation result of reference genome independence;Integrate structural transcript annotation information, filter transcript that cannot be compared with reference genome, obtain new transcript annotation information;Integrate sample high-throughput transcriptome data, and the pan-transcriptome annotation information of target eukaryotic species can be obtained.The present application can obtain more complete transcriptome annotation structure of eukaryote, and different strategies are mutually verified, which greatly improves the annotation accuracy.

An engineered tyrosyl-tRNA synthetase / tRNA pair that co-translationally incorporates O-sulfotyrosine in response to UAG codons in E. coli and mammalian cells is described herein. This platform enables recombinant expression of eukaryotic proteins homogeneously sulfated at chosen sites.

The invention relates to a modular culture system for growing prokaryotic and / or eukaryotic organisms using a nutrient solution (N) for the organisms, wherein the culture system comprises a series of culture modules (100) for receiving the organisms, which culture modules are interconnected to conduct the nutrient solution (N), and an extraction module (200), connected to one of the culture modules (100) for conducting the nutrient solution (N), for the controlled extraction of the nutrient solution (N) from the culture module (100).Each culture module (100) comprises: an outer container (110) through which the nutrient solution (N) flows in a flow direction (S); an inner container (120), located at least partially within the outer container (110), for receiving the organisms; and several connection devices (130) for connecting the outer container (110) to the outer container (110) of an additional culture module (100) and / or to the extraction module (200) for conveying the nutrient solution (N). The inner container (120) is permeable to the nutrient solution (N), and a nutrient chamber (115), through which the nutrient solution (N) flows, is located between the outer container (110) and the inner container (120). The extraction module (200) comprises an extraction control device (240) for controlling the flow of the nutrient solution (N) extracted from the cultivation module (100) by means of the extraction module (200).

The invention discloses a CAR (chimeric antigen receptor) recombinant lentiviral expression vector suitable for CIK (cytokine-induced killer) cells and NK (natural killer) cells to express EpCAM antibodies, IL-21 and CCL19, and belongs to the technical field of genetic engineering and biology. According to the expression vector, a CAR (chimeric antigen receptor) gene structure, an IL-21 (interleukin-21) gene expression structure and a CCL19 (chimeric antigen receptor 19) gene expression structure are connected between multiple cloning sites Nhe1 and Not1 of a vector pCHD-CMV-MCS-EF1-copGFP-T2A-puro, so that a recombinant plasmid pCDH-21 * 19 EpCAM-CAR is obtained. And transforming the recombinant plasmid into escherichia coli DH5alpha to obtain the recombinant microbial cell. The pCDH-21 * 19 EpCAM-CAR recombinant lentiviral expression vector is packaged, lentiviral particles are concentrated, CIK cells and NK cells are infected, and eukaryote transgenic cells are obtained. The CAR vector aiming at the solid tumor colorectal cancer is constructed, the design of the vector aims at improving the activating and killing capacity of CIK cells and NK cells and solving the problems of tumor microenvironment immunosuppression and the like, and a foundation is further laid for the research of treating solid tumors by applying a CAR technology.

The present invention discloses a novel genome editing tool EbCas12a-D141R and its application in gene editing, belonging to the field of biotechnology. The EbCas12a-D141R is a variant of the type V class II CRISPR protein EbCas12a, and its amino acid sequence is as shown in SED ID NO.1 in prokaryotic cells and as shown in SED ID NO.2 in eukaryotic cells. The present invention for the first time obtains a variant EbCas12a-D141R with higher efficiency based on the type V class II CRISPR protein EbCas12a (1158AA), and the gene editing efficiency of this variant in eukaryotic cells is significantly higher than that of EbCas12a. The discovery of EbCas12a-D141R further expands the types of gene editing tools and has a very important role in basic scientific research and clinical treatment.

The present invention relates to minimal polypeptide-encoding RNA molecules, in particular, RNA molecules that lack or comprise shortened regulatory sequences as compared to those found in eukaryotic mRNAs, which are thus more amenable to chemical synthesis. The invention also encompasses compositions comprising distinct populations of said RNA molecules, each encoding a unique polypeptide, and methods for manufacturing and using the RNA molecules or compositions.

The application discloses an ELISA kit for detecting anti-chromatin antibodies. The kit contains purified chromatin monomers with stripped H1 histone components in a natural structure. The application also discloses a detection method for the ELISA kit. The chromatin structure in the eukaryotic cell nucleus is complex, and is a natural complex composed of 40% DNA, 40% histone, 20% non-histone, a small amount of RNA and other macromolecules. The purification technology is high in requirement, and the price of the natural purified chromatin monomers is high. The application purifies the chromatin monomers with stripped H1 histone components according to the structure of the natural chromatin, retains the effective antigen components, and greatly reduces the cost. The kit can be used for detecting the anti-chromatin antibodies, fills the blank of the domestic anti-chromatin antibody detection method, and has the characteristics of large detection flux, accurate result, high sensitivity and simple operation.

The application provides an RNA framework for gene editing and a gene editing method, the RNA framework comprising a sequence upstream of a target site, a sequence to be inserted, and a sequence downstream of the target site in the 5' to 3' direction. The gene editing method is based on the inherent mechanism of eukaryotes, uses RNP or RNA (which can be prepared in vitro) and related proteins as carriers, and is transferred into the cytoplasm and nucleus to realize gene editing on a specific sequence or site on the genome of a target system, such as insertion, deletion, replacement and site replacement of a specific sequence, and has high targeting accuracy. The application is more suitable for further clinical application than other prior art because no exogenous system or substance such as protein derived from prokaryotes is introduced, and no double-strand break is generated.

The invention relates to the technical field of kits, and discloses a peanut allergen component fluorescence PCR (Polymerase Chain Reaction) detection equivalent kit, which comprises (1) 2 * Probe qPCR Mix; (2) a florescence source probe primer mixture; (3) a 18S internal reference probe primer mixture; (4) ultrapure water; (5) positive control; (6) negative control; according to the invention, a dual detection mechanism of a florescence source specific gene and an eukaryote general reference gene is combined, and a specially designed chimeric recombinant plasmid is adopted as a multifunctional positive control, so that when a complex food matrix is treated, a forced standard reaching mechanism of a reference gene channel can timely identify a sample treatment abnormal condition; and wrong interpretation caused by improper operation is avoided. Meanwhile, the activity of core components of the kit can be verified through the two-channel synchronous response characteristic of positive control, and the credibility of a detection result and the scientific preciseness of interpretation are improved.