39results about "Hydrolases" patented technology

Methods and compositions for producing siRNAs, e.g., in the form of a d-siRNA composition, from dsRNAs are provided. In the subject methods, a dsRNA is contacted with a composition that includes an activity that cleaves dsRNA into siRNAs, where the composition efficiently cleaves dsRNA into siRNAs. siRNAs produced by the subject methods find use in a variety of applications, particularly in applications where the specific reduction or silencing of a gene is desired. Also provided are kits for use in practicing the subject invention.

The invention provides a genome editing method based on a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) system and belongs to the technical field of gene editing. The genome editing method provided by the invention utilizes introns to express guide RNA molecules in the CRISPR genome editing system and adopts a principle that one or more than one tRNA-gRNA or crRNA series unitsare arranged in the introns of an encoded gene, and after the introns form a fusion gene with Cas9 or Cpf1, a promoter can be used for driving. The genome editing method provided by the invention hasthe benefits that a cellular endogenous mRNA splicing system and a cellular endogenous tRNA processing system are skillfully utilized, and other elements are not required to be added, so that not onlyis the safety higher, but also more simplicity can be realized; the synchronous expression of a plurality of guide RNAs with the Cas9 or the Cpf1 is realized by utilizing the one promoter, so that anexisting CRISPR editing system is simplified, moreover, the efficiency and the ability of simultaneously editing multiple target points of the CRISPR editing system are improved.

The invention is directed to transcription activator-like effector nuclease (TALEN)-mediated DNA editing of disease-causing mutations in the context of the human genome and human cells to treat patients with compromised genetic disorders.

It is an object of the present invention to provide a solution for creating an effective method for retarding unhealthy manifestations brought by ageing of human beings (in particular, but not limited to the reduction of sexual activity and fertility, climax, changes in glucose tolerance, reduction of cognitive and mnestic functions, reduction of stress resistance, development of organ and tissue sclerosis) without directly affecting the genetic apparatus of the ageing cells.

According to the invention this task is solved by administration into the blood circulation of the agent which inactivates extracellular blood plasma DNA; the extracellular blood DNA inactivating agent can be embodied in the form of an extracellular blood plasma DNA destroying agent; said extracellular blood plasma DNA destroying agent can be embodied in the form of an DNase enzyme; the extracellular blood plasma DNA inactivating agent can also be embodied in the form of an extracellular blood plasma DNA binding agent; the extracellular blood plasma DNA binding agent can be embodied in the form of anti-DNA antibodies; the extracellular blood plasma DNA inactivating agent can be administered in the form of an enzyme modifying the chemical composition of extracellular blood plasma DNA; the extracellular blood plasma DNA inactivating agent can be embodied in the form of an agent that stimulates synthesis or activity of endogenous deoxyribonuclease, or an agent that stimulates the synthesis of antibodies which capable to bind extracellular blood plasma DNA.

The invention provides a clone and a pronucleus expression method of a novel B class epoxide hydrolase gene mature peptide cDNA sequence from Aspergillus usamii E001. The nucleotide sequence of the novel B class epoxide hydrolase gene mature peptide cDNA sequence is SEQ ID NO:1, the corresponding amino acid sequence is SEQ ID NO:2, and the corresponding gene is named Aus EH-B. According to the invention, good stereoselectivity is achieved for (R)-epichlorohydrin through chiral gas chromatography analysis rEH, and a produced (S)-epichlorohydrin antipode excess value achieves 99%. The pronucleus expression method provided by the invention lays the foundation for the industrialized production of epoxide hydrolase and provides the basis for preparing chiral epichlorohydrin by researching an EH enzyme kinetic resolution method for biological catalysis technology industrialization.

The invention belongs to the field of microbial genetic engineering, and discloses a GTP cyclohydrolase I gene folE and application. A nucleotide sequence of the GTP cyclohydrolase I gene folE is shown as SEQ ID NO:1. The gene is derived from food-borne lactobacillus plantarum, is safe and can be used in the field of later-stage food fermentation. A gene knockout strain screening method can be used for improving the screening efficiency of knockout strains. The key role of gene folE gene in folic acid synthesis is proved, and a certain theoretical basis is provided for the research and development of folic acid synthesis functional food. The GTP cyclohydrolase I gene folE and application analyze that the gene is derived from food-grade microorganisms and is safe; and the GTP cyclohydrolaseI gene folE and application analyze that the gene folE gene has nothing to do with cell morphology and strain growth except playing an important role in folic acid synthesis.

A recombinant vector for delivering A3G genes into human cells comprising (i) a gene expression block including an A3G gene selected from a wild type A3G gene represented by SEQ ID NO: 1 and a mutant A3G gene and (ii) a group of elements from a modified lentiviral vector including lentiviral regions of packaging signal (ψ, psi), LTRs, RRE, and PBS; wherein said A3G gene is operably linked to the packaging signal (ψ, psi), LTRs, RRE, and PBS.

Compositions and methods for protecting a plant from an insect pest are provided. In particular, nucleic acid sequences encoding insect protoxins modified to comprise at least one proteolytic activation site that is sensitive to a plant protease or an insect gut protease are provided. Cleavage of the modified protoxin at the proteolytic activation site by a protease produces an active insect toxin. Methods of using the modified insect protoxin nucleic acid sequences and the polypeptides they encode to protect a plant from an insect pest are provided. Particular embodiments of the invention further provide modified insect protoxin compositions and formulations, expression cassettes, and transformed plants, plant cells, and seeds.

The present invention relates to methods for obtaining transgenic embryos, animals, and plants. Also encompassed in the present invention are transposase-mediated trangenesis methods including transposase-mediated intracytoplasmic sperm injection (TN:ICSI), transposase-mediated intracytoplasmic round spermatid injection (TN:ROSI), and transposase-mediated in vitro fertilization (TN:IVF).

It is intended to provide an extract, kit and a process for synthesizing a protein in a cell-free system. The extract is characterized by being prepared from mutant Escherichia coli cells having a mutation in the ribosomal protein S12 gene. In a preferred embodiment, the mutation is such a mutation as conferring a resistance or dependence to streptomycin on E. coli whereby the reading efficiency of mRNA codon on ribosomes can be improved and thus the protein productivity can be elevated.

The invention discloses Cas9 fused protein capable of improving the gene knock in efficiency and a safe efficient exogenous gene knock in and integration system. According to the Cas9 fused protein capable of improving the gene knock in efficiency and the safe efficient exogenous gene knock in and integration system, by fusing the Cas9 protein and DNA binding domains of a transcription factor to produce the Cas9 fused protein for gene knock in, the gene knock in efficiency can be effectively improved. According to the exogenous gene knock in and integration system, on the basis of applying theCas9 fused protein, a small-molecule compound is further used for intervening cells co-transfected by an expression vector, which empresses the Cas9 fused protein and gRNA, and a donor vector, and bymeans of a synergistic effect of the Cas9 fused protein and the small-molecule compound, occurrence of KI events of different genotypes and cell lines can be effectively promoted, so that the gene knock in efficiency is significantly improved, and convenience is provided for production of model animals and gene therapy.

Described herein are RNA-based antisense therapeutics to target antibiotic resistant bacteria. The antisense therapeutics disclosed herein can be useful in re-sensitizing drug-resistant bacteria to antibiotics, as well as developing antibiotics that have bactericidal or bacteriostatic effects on the drug-resistant bacteria or are capable of preventing emergence of antibiotic resistance. Also described herein are methods for re-sensitizing a subject to one or more antibiotics in need thereof, methods for identifying target genes involved in adaptive antibiotic resistance in bacteria, and methods for developing antibacterial antisense therapeutics.

The invention relates to recombinant plasmid pDSBCm harboring the gene vapk-repeated region, which gene encodes alkalic protease VapK, a microorganism Vibrio metschnikovii transformed therewith, and method for producing an alkaline protease VapK using the same microorganism.

The title of the invention is polynucleotide adapter design for reduced bias. Compositions and methods of use are provided that among other things, allow for efficient adapter ligation to small RNAs.Embodiments of the compositions include partially double stranded polynucleotides for use as 3' adapters that contain a cleavable linker positioned between a single-stranded region and a doublestranded region. Upon ligating the 3' adapters, the single-stranded region is released by cleaving the cleavable linker.

A method obtains a biological glue from a bacteria.

The invention discloses a low-temperature lipase Lip1 and a gene thereof. An amino acid sequence of the low-temperature lipase is SEQ ID NO:2; and the gene is from Yersinia enterocolitica KM1 of a Kunming slaughter house refrigeration house. According to the invention, the low-temperature lipase is prepared from a recombinant expression vector containing the enzyme gene is used for preparing low-temperature lipase; the low-temperature lipase prepared from the expression vector has good activity and stability under the low-temperature condition, wherein the optimal acting temperature is 25 DEG C and the optimal acting pH value is 7.5, and therefore, the low-temperature lipase can be extensively applied to the industrial fields including detergent preparation and biodiesel preparation.

The present invention relates to engineered carbohydrate-active enzyme constructs that are useful for glycan polymers synthesis. The construct comprises a CD domain from a GH, wherein the domain is conjugated to CBM3a via a peptidic linker. The invention also relates to a method of improving glycan polymer synthesis by using engineered carbohydrate active enzymes comprising a CD domain and CBM3a.

Disclosed are methods of altering expression of a gene with a promoter region CTCF binding site. Also disclosed are compositions and methods useful for treating a disease or condition involving over-expression or under-expression of a gene with a promoter region CTCF binding site. Further disclosed are cells and non-human animals with modified a promoter region CTCF binding site, as well as methods for screening for compounds that can modify the expression of a gene with a promoter region CTCF binding site.

The invention discloses an upland cotton phosphatidylinositol specific phospholipase C gene GhPIPLC2-2. The nucleotide sequence of cDNA (complementary deoxyribonucleic acid) of the gene is shown in SEQ ID No.1 in the description, and an amino acid sequence encoded by the gene is shown in SEQ ID No.2 in the description. The invention further discloses application of the gene GhPIPLC2-2 in culturingsalt-resistant plants or improving salt resistance of plants. Experiments show that arabidopsis with overexpression of the GhPIPLC2-2 gene is sensitive to salt, the salt resistance of a plant can beremarkably improved through gene knockoff or RNAi (ribonucleic acid interfere), and it indicates that the GhPIPLC2-2 gene has significant functions in application in culturing salt resistant plants.

The present invention belongs to the field of biotechnology. More specifically, the present invention provides a protease, a non-naturally occurring fusion protein comprising a corresponding protease recognition site, expression vectors encoding same, host cells comprising said expression vectors, kit of parts as well as methods applying the protease, fusion protein, and uses thereof, as defined in the claims. The presently disclosed protease/protease recognition site is particularly useful in methods requiring an orthogonal set of proteases, and is suitable for use in both prokaryotic and selected eukaryotic expression systems