686 results about "Gene Modification" patented technology

The invention relates to a gene modified cell and a method for producing fucosyllactose by using the gene modified cell, in particular to a saccharomyces cerevisiae gene modified cell and application thereof, and belongs to the technical field of biological gene engineering. The present invention provides a genetically modified cell comprising a recombinant nucleic acid sequence encoding a polypeptide having an [alpha]-1, 2-fucosyltransferase polypeptide ([alpha]-1, 2-fucosyltransferase) activity. The alpha-1, 2-fucosyltransferase polypeptide includes, but is not limited to, polypeptides having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical amino acid sequences with the amino acid sequences of SEQ ID NO: 1-25. According to the gene modified cell, the fucosyllactose synthesis capability of an original strain is improved.

The invention relates to a gene modified cell and a method for producing fucosyllactose by using the gene modified cell, in particular to a kluyveromyces kluyveromyces gene modified cell and application thereof, and belongs to the technical field of biological gene engineering. The present invention provides a genetically modified cell comprising a recombinant nucleic acid sequence encoding a polypeptide having an [alpha]-1, 2-fucosyltransferase polypeptide ([alpha]-1, 2-fucosyltransferase) activity. The alpha-1, 2-fucosyltransferase polypeptide includes, but is not limited to, polypeptides having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical amino acid sequences with the amino acid sequences of SEQ ID NO: 1-25. According to the gene modified cell, the fucosyllactose synthesis capability of an original strain is improved.

Systems and methods for targeted gene modification, targeted insertion, perturbation of gene transcripts, and nucleic acid editing. Novel nucleic acid targeting systems comprise components of CRISPR systems and non-LTR retrotransposon elements.

The present invention relates to ARP2 of plant stigma cells; the invention relates to application of Ca < 2 + > transport capacity damage of homologous genes 1 and 1 in breaking through reproduction isolation of plants. According to the method for breaking through the plant reproductive isolation, a foundation is laid for further research on the relationship of small peptides for regulating and controlling reproductive isolation calcium signals through co-receptors, and a simple and effective target is provided for gene modification for breaking through the reproductive isolation.

The invention discloses bacillus subtilis with high yield of surfactin, a related enzyme mutant and a preparation method of the bacillus subtilis, the bacillus subtilis is obtained by a gene modification method and can produce the surfactin with high yield, and the highest yield can reach 9.4 g / L; the invention also provides key enzyme mutants participating in surfactin synthesis in the bacillus subtilis, and the key enzyme mutants comprise a biotin carboxylase IdeHA mutant, a biotin carboxylase accBC mutant, an enoyl reductase FabI mutant, a malonyl transferase FabD mutant and a dihydrolipoyl dehydrogenase IPdU mutant. The key enzyme mutants comprise a biotin carboxylase IdeHA mutant, a biotin carboxylase accBC mutant, an enoyl reductase FabI mutant, a malonyl transferase FabD mutant and a dihydrolipoyl dehydrogenase IPdU mutant. The bacillus subtilis for high-yield surfactin and the preparation method thereof disclosed by the invention are beneficial to realizing industrial production and commercial application of the surfactin.

The invention belongs to the technical field of stem cell culture and exosome extraction, and provides an RELM beta pre-activated stem cell outer vesicle with high anti-inflammatory and anti-fibrosis activity. According to the method, the mesenchymal stem cells are subjected to hunger culture after being adhered to the wall and then are continuously cultured by a complete culture medium containing RELM beta factors, then supernate is collected, and the exovesicles are obtained through separation. In the preparation process of the extracellular vesicles, the paracrine function of MSC cells is promoted by the RELM beta, the secretion of anti-inflammatory and anti-fibrosis active factors is obviously activated, and the generation of the extracellular vesicles with enhanced anti-inflammatory and anti-fibrosis effects is promoted. Gene modification is not involved, toxic biochemical reagents or materials are not added, exogenous operation steps are simple and convenient, clinical application is guaranteed through sterile operation, no RELM beta residues exist after extracellular vesicle extraction, and the method is safe, non-toxic and effective.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.

In one aspect, the disclosure relates to a gene modification tool that fuses a Cas protein with a polymerase. This DSB-independent approach utilizes a DNA-RNA chimeric prime-editor-like guide RNAs (chimeric pegRNAs or cpegRNAs) to enable programmable DNA sequence replacement or excision at endogenous human genomic sites. The Cas9 and RNA portion of cpegRNA helps with targeting a gene, while the DNA portion of cpegRNA is used as a template by the polymerase to repair. Furthermore, the disclosed method can be used to introduce large DNA modifications in the genome.

The invention belongs to the technical field of biology, and particularly relates to a genetically modified mesenchymal stem cell as well as a preparation method and application thereof. The invention relates to a gene modified mesenchymal stem cell, the gene modified mesenchymal stem cell is a mesenchymal stem cell of an over-expression SIRT6 gene, and the acquisition number of the SIRT6 on NCBI (National Center of Biotechnology Information) is NM001193285. The invention finds that SIRT6 is helpful for delaying the cell senescence process and obviously improving the anti-senescence capability of cells by regulating senescence-related genes and proteins.

Provided herein are compositions and methods for use in challenging pathogenic bacteria on plants. Optional features include modification of a donor bacteria to include exogenous nucleic acids encoding for conjugation machinery and gene modification components, such as guide sequence for use in CRISPR. The compositions and methods provided herein can be used for delivery to a wide variety of crops and for targeting one or more pathogens.

The invention relates to a Kluyveromyces cerevisiae gene modified cell containing GDP-mannose dehydratase polypeptide, which can be applied to synthesis of human milk oligosaccharide, and belongs to the field of synthetic biology. According to the technical scheme, the gene modified cell comprises a recombinant nucleic acid sequence for coding a polypeptide with GDP-mannose dehydratase polypeptide (GDP-mannose dehydratase polypeptide) activity. The polypeptide capable of catalyzing the conversion of GDP-mannose into GDP-4-keto-6-deoxymannose comprises a polypeptide of which the amino acid sequence is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2 and / or the amino acid sequence of polypeptides M1-M13, wherein the amino acid sequence of the polypeptide is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of the polypeptides M1-M13. The invention provides a kluyveromyces kluyveromyces genetically modified cell with fucosyllactose synthesis activity.

The invention relates to a genetically modified cell, and belongs to the technical field of biological genetic engineering. The invention provides a genetically modified cell, which contains a heterologous nucleic acid sequence of an alpha-1, 3-fucosyltransferase polypeptide. The alpha-1, 3-fucosyltransferase polypeptide further comprises but is not limited to derived polypeptides M1-M21 of the polypeptide with the amino acid sequence as shown in SEQ ID NO: 1, and compared with the polypeptide as shown in SEQ ID NO: 1, the 3-FL catalytic synthesis capability of the derived polypeptides and the specificity of a catalytic substrate are both improved.

The invention discloses a production method of fucosylated oligosaccharide. The core of the method is as follows: the GDP-D-rhamnose is converted into the GDP-L-fucose by using the GDP-D-rhamnose-3, 5-epimerase; the GDP-D-rhamnose is obtained by converting the GDP-D-mannose-4, 6-dehydratase and the GDP-4-keto-6-deoxy D-mannose reductase by taking the GDP-D-mannose as a substrate, and the GDP-D-rhamnose is obtained by taking the GDP-D-mannose as a substrate. The invention relates to a method for producing a fucosylated oligosaccharide, which is characterized in that a GDP-D-mannose-4, 6-dehydratase, a GDP-4-keto-6-deoxyD-mannose reductase, a GDP-D-rhamnose-3, 5-epimerase and a fucosyltransferase are expressed in a genetically modified host cell, and the fucosylated oligosaccharide can be produced by the way mentioned above.

A composition for genetic editing of a hydroxyacid oxidase 1 (HAO1) gene, comprising: (a) a messenger RNA (mRNA) encoding a Cas12i2 polypeptide, (b) a guide RNA (gRNA) targeting the HAO1 gene; and (c) lipid excipients, which optionally form lipid nanoparticles. Also provided herein are methods for genetic modification of the HAO1 gene in host cells with the composition provided herein.

The present disclosure provides engineered bacterial cells having one or more genetic modifications that result in increased production of expression products, such as increased production of plasmids with reduced occurrence of plasmid concatemers or multimers. Also disclosed herein are expression systems for product manufacturing in media having reduced antibiotic concentration, as well as methods of making cells for use in the expression systems. The cells and expression systems increase the growth rate and production yields of biomolecules produced within the host bacterial host cells without the use of antibiotics, and can produce the product with little to no endotoxins present.

The invention discloses a construction method and application of a chlamydomonas reinhardtii-sourced high-sweetness protein variant engineering algal strain, the construction method comprises the following steps: carrying out gene modification on sweeteners Thaumatin and Brazzein, optimizing preference codons of chlamydomonas reinhardtii to synthesize CrThaumatin and CrBrazzein encoding genes, cloning the encoding genes into an expression vector pGM6, introducing the recombinant expression vector into a chlamydomonas reinhardtii wild-type chlamydomonas sp. Algal strain, and carrying out high-sweetness protein variant engineering algal strain of the chlamydomonas reinhardtii-sourced high-sweetness protein variant engineering algal strain of the chlamydomonas reinhardtii. And screening by using a paromomycin resistance marker carried by the expression vector to successfully obtain an engineering algal strain. According to the present invention, the engineering strain is subjected to fermentation production to obtain the dry and stable Chlamydomonas reinhardtii powder rich in the target sweet protein, and the protein purification oral test results show that the sweet taste of the Chlamydomonas reinhardtii powder is 3-5 times of the sweet taste of the natural Thaumatin and Brazzein protein;

Provided herein include methods and compositions for effecting a targeted genetic change in DNA in a cell. Certain aspects and embodiments relate to improving the efficiency of the targeting of modifications to specific locations in genomic or other nucleotide sequences. As described herein, nucleic acids which direct specific changes to the genome may be combined with various approaches to enhance the availability of components of the natural repair systems present in the cells being targeted for modification.

Provided herein are methods and compositions for genetically engineering a cell (e.g., a hematopoietic cell) using CRISPR / Cas systems and homology-directed repair, genetically engineered cells produced by such methods, and methods involving administering such genetically engineered cells to a subject, such as a subject having a genetic disease.

The invention provides an anti-tumor modification method of stem / progenitor cell-derived neutrophil, which comprises the following steps: extracting mouse bone marrow-derived Lin-progenitor cells and human umbilical cord hematopoietic stem / progenitor cells, modifying ER-Hoxb8 progenitor cells by gene editing, and inducing the ER-Hoxb8 progenitor cells into neutrophil; three modification strategies of chimeric antigen receptor CAR gene engineering modification, high expression MET gene modification and high expression of IFN-gamma while the TGFBR2 gene is stably knocked out are respectively adopted, and the anti-tumor phenotype of the neutrophil N1 is maintained. Compared with the prior art, the engineering neutrophile granulocyte has the following advantages that the engineering neutrophile granulocyte prepared by the method has the advantages of infinite sources, long-time preservation, safety, controllability, efficient and rapid preparation and the like; the engineered neutrophil can maintain the N1 antitumor phenotype and promote the antitumor function; the immunosuppression microenvironment of the solid tumor can be improved and the treatment effect can be improved by singly using or combining the immune checkpoint inhibitor.

The invention provides to improved methods for the modification of genes in plant cells, and plants and seeds derived therefrom. More specifically, the invention relates to the increased efficiency of targeted gene mutation by combining gene repair oligonucleotides with approaches that enhance the availability of components of the target cell gene repair mechanisms.

The invention relates to an IL5RA cell surface marker. A cell tag comprising an extracellular region, a transmembrane region, and optionally an intracellular region is provided. The extracellular region comprises an IL5 receptor alpha (IL5Ra) sequence linked to a transmembrane domain, and the recombinant polypeptide cannot function in signal transduction. The cell tag can be operably linked to a transgene. Expression of the cell tag allows for the identification, detection, selection, and ablation of cells expressing the transgene and the cell tag. In some embodiments, the genetically modified host cell comprises a transgene comprising a polynucleotide encoding a chimeric antigen receptor comprising a ligand binding domain and a polynucleotide encoding a cell tag. Pharmaceutical formulations produced by the methods and methods of using the pharmaceutical formulations are also described.

The disclosure relates to compositions, systems, and methods related to repair RNA sequences, as well as related systems for trans-splicing a target nucleic acid using a repair RNA sequence. Various compositions and systems are described herein, including compositions comprising a repair RNA sequence comprising one or more exons and / or introns, a splice donor and / or splice acceptor, wherein the repRNA is suitable for trans-splicing. The compositions, systems, and methods described herein are suitable for targeting one or more Usher syndrome-associated genes, and treating Usher syndrome.

The present disclosure provides genetically modified iPSC-derived γδT cells and their precursors. A double genomic disruption in the suppressor of cytokine signaling 1 (SOCS1) gene and the cytokine-inducible sh2-containing protein (CISH) gene are provided, as is a triple genomic disruption in genes for SOCS1, CISH, and Bcl-2 interacting mediator of cell death (BIM), as is a quadruple genomic disruption in genes for SOCS1, CISH, BIM, and cell surface death receptor (FAS), as is a quintuple genomic disruption in genes for SOCS1, CISH, BIM, β-2-Microglobulin (B2M), and class II transactivator (CITTA), as is a sextuple genomic disruption in genes for SOCS1, CISH, BIM, B2M, CITTA, and FAS. Also provided is genetically modified iPSC-derived γδT cells and their precursors with improved proliferation and tumor killing activity. Also provided are genetically modified iPSC-derived γδT cells and their precursors further comprising CD19 CAR. The present disclosure further provides methods making and using such cells, as well as gene editing systems.

The invention discloses a tryptophan synthetase trp B modified gene and a preparation method of 6-fluoro-L-tryptophan, belongs to the technical field of gene modification and amino acid synthesis, and provides a tryptophan synthetase trp B gene subjected to mutation modification, and the nucleotide sequence of the tryptophan synthetase trp B gene is shown as SEQ ID NO.1. After gene modification, substrate molecules can enter the channel more easily. After gene modification, a channel of an active center of tryptophan synthetase is more beneficial to entering of substrates 6-fluoroindole and L-serine, and the yield of 6-fluoro-L-tryptophan is increased. According to the method, a specific strain containing tryptophan synthetase is adopted, tryptophan synthetase can be efficiently expressed by culturing in an optimized culture medium, so that the 6-fluoro-L-tryptophan synthesized by an enzymic method has relatively high catalytic rate and conversion rate, the molar conversion rate of L-serine reaches 86.5% or above, the reaction conditions of the enzymic method synthesis are mild, the enzyme stereoselectivity is strong, the catalytic efficiency is high, and the method is suitable for industrial production. The method is suitable for industrial production.

The present invention belongs to the field of bioengineering, genetic engineering technology and oligosaccharide (rare sugar) synthesis technology, and specifically relates to a method for producing lactose-N-disaccharide using genetically modified recombinant yeast. The technical scheme of the present invention is: a recombinant yeast cell is introduced with (1) a heterologous lactose-N-disaccharide phosphorylase ( LNBP ) gene, and (2) fructose-1-phosphate phosphorylase ( YQAB ) gene, and further preferably, also includes (3) destruction or modification of a marker gene. The recombinant yeast cell has the ability to synthesize lactose-N-biose (Lacto-N-biose, LNB) from lactose, and the yield of LNB is greatly improved by a molecular switch.

The invention relates to pseudomonas putida engineering bacteria for producing rhamnolipid as well as a construction method and application of the pseudomonas putida engineering bacteria, the engineering bacteria take pseudomonas putida KT2440 with phaC1, phaC2, algD and fliC gene deletion as a chassis cell, and rhlAB, rmlACBD, algC and FabY genes are over-expressed in the chassis cell. Pseudomonas putida KT2440 with biological safety is used as a chassis cell for gene modification, the constructed engineering bacteria are used for fermentation production of rhamnolipid, on the basis of rhamnolipid yield increase, good physiological adaptability and fermentation stability are shown, and the rhamnolipid yield is increased. And the modified engineering strain can stably and efficiently produce rhamnolipid with a specific structure.

The invention discloses a method for improving the heterologous soluble expression efficiency of cytochrome P450, which is characterized in that plasmids containing coding genes of fungus-derived cytochrome P450 truncated protein and plasmids containing molecular chaperones are jointly transferred into host bacteria to obtain recombinant genetically engineered bacteria for improving the heterologous soluble expression efficiency of cytochrome P450. Through host screening, molecular chaperonin and cytochrome P450 gene modification, the soluble expression efficiency of cytochrome P450 in Escherichia coli is improved, and the problems of poor protein solubility and low expression efficiency when CYP450 is expressed in Escherichia coli at present are solved.

The invention discloses bacillus subtilis with high yield of specific surfactin and a preparation method and application thereof, the bacillus subtilis is obtained by a gene modification method and can produce high yield of C15 component surfactin, and the highest yield can reach 72.5%; the invention also provides an enzyme mutant participating in the production of the surfactin from the bacillus subtilis, wherein the enzyme mutant comprises carboxyl transferase accAD, biotin carboxylase IdeHA, biotin carboxylase accBC and fatty acid peroxygenase cypC; in addition, the invention also provides a preparation method and application of the bacillus subtilis. The bacillus subtilis for high-yield production of specific surfactin and the preparation method thereof provided by the invention are beneficial to increase of the yield of surfactin, especially industrial production and commercial application of C15 component surfactin.

The invention relates to a genetically modified licorice stem cell and a preparation method thereof, and relates to the technical field of genetic engineering, and the licorice stem cell is modified by introducing a gene coding dimethyl allyl transferase (PcM4DT) or an active fragment thereof and a gene coding isoflavone synthase (IFS) or an active fragment thereof into the licorice stem cell. The content of secondary metabolites in the genetically modified liquorice stem cells is remarkably increased compared with that of unmodified liquorice stem cells, the increased secondary metabolites meet the industrial production requirement and can be directly used for high-end skin care products and medical products, and the liquorice resource value is expanded.

The present invention provides a non-human animal expressing a human or chimeric (e.g., humanized) KLRG1 protein and methods of use thereof.