585 results about "Gene Modification" patented technology

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.

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.

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 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 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 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 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 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 invention relates to the technical field of adhesives, in particular to a cottonseed protein-based environment-friendly adhesive for plywood and a preparation method of the cottonseed protein-based environment-friendly adhesive. The cottonseed protein-based environment-friendly adhesive for plywood is composed of gene modified cottonseed protein isolate, double modified starch, an intelligent cross-linking agent system, a reinforced fiber system and the like. The cottonseed protein is subjected to gene editing and hydrophilic modification, and the starch is grafted with the nano silicon dioxide through esterification; the cross-linking agent contains a temperature-sensitive material and a dynamic cross-linking unit, and the reinforced fibers are nanocellulose and graphene oxide compounds; various functional auxiliaries are supplemented, and all the components have a synergistic effect, so that the high-performance environment-friendly adhesive is formed. The adhesive is mainly prepared from natural raw materials and is free of formaldehyde release; through gene modification, intelligent crosslinking and other technologies, the fluidity, the bonding strength and the water resistance are improved, and the bonding strength after boiling in boiling water is larger than or equal to 0.7 MPa; storage is stable, the process is simple, cost is low, and green development of the plywood industry is promoted.

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 (CIITA), as is a sextuple genomic disruption in genes for SOCS1, CISH, BIM, B2M, CIITA, 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.

Provided herein are modified nucleic acids comprising modified MTM1 genes, modified muscle-specific promoters, e.g., MHCK7, or a combination thereof. Also provided herein are expression cassettes, AAV vector genomes, and AAV particles comprising the same. Further provided herein are methods of delivering MTM1 to a muscle cell and methods of treating XLMTM.

The invention provides a recombinant oncolytic virus for treating solid tumors with rare and rare gene mutations, which is an OAV treatment platform subjected to triple virus structural protein gene modification and triple adenovirus serotype chimerism, and is named as NeoViron. NeoViron can directly deliver tumor neoantigens to a plurality of solid tumors, especially tumors with rare mutations, and a new general strategy is provided for treating a plurality of intractable tumors.

The invention relates to a gene modified cell for producing HMOs, which can be applied to synthesis of difucosyllactose, and belongs to the technical field of gene engineering. The invention provides a genetically modified cell, which contains an alpha-1, 2-fucosyltransferase polypeptide and a heterologous nucleic acid sequence of the alpha-1, 3-fucosyltransferase polypeptide. The genetically modified cell has higher activity of synthesizing difucosyllactose, and does not contain synthetic 3-fucosyllactose or lower side activity of synthetic 2 '-fucosyllactose. The technical scheme of the invention has positive significance for industrial production of human milk oligosaccharide, and the method is green, efficient and sustainable, is beneficial to industrial large-scale production, and has important practical value.

The present invention relates to a method for obtaining a genetically modified progeny of an avian animal by packaging a gene edited or genetically modified vector in a recombinant adenovirus and injecting into the dorsal aorta of an embryo when PGCs migrate to a developing gonad; compared with the existing disclosed method for transforming the PGCs in vitro, the operation method disclosed by the invention not only can stably and effectively infect the PGCs in vivo, but also has the advantages that more importantly, the probability of generating gene editing offspring is higher, and particularly, the breeding efficiency of cultivating the transgenic chicken offspring with KRT75L4 gene and MSTN gene knockout is higher; and on the other hand, the reporter gene or crispr-cas protein gene does not need to be integrated into the genome of the bird and the non-native protein does not need to be expressed, so that the operability is higher from the commercial level.

The present disclosure relates to compositions and methods for modifying target nucleic acids, and methods for detecting nucleic acids. Various compositions are described herein, including compositions comprising an endonuclease, an endonuclease system, and a chimeric protein having the endonuclease and a nucleic acid modulating domain or a nucleic acid modifying domain.

A genetically modified yeast cell, wherein the yeast cell comprises a genetic modification comprising overexpression of yeast gene encoding porphobilinogen deaminase (HEM3), the HEM3 gene having at least 80% identity with SEQ ID No. 7. The genome of the modified yeast cell further comprises one or more genetic modifications in one or more genes selected from: genes coding for heme-dependent repressor of hypoxic genes (ROX1), genes coding for heme oxygenase (HMX1), genes coding for a receptor for vacuolar proteases (VPS10), and genes coding for vacuolar proteinase (PEP4), the one or more genetic modifications being such that expression of a polypeptide from such a gene is reduced or disrupted or the polypeptide expressed is non-functional.

The application relates to a T cell-targeting nanoparticle based on a nucleic acid aptamer and a preparation method and application thereof, in particular to a T cell-targeting lipid aptamer, wherein the lipid aptamer comprises a hydrophobic molecule and a nucleic acid aptamer, the nucleic acid aptamer comprises at least one nucleic acid aptamer for targeting a T cell surface molecule, and the molar ratio of the hydrophobic molecule and the nucleic acid aptamer is 5000:1-1:10. The lipid aptamer or the nanoparticle formed by self-assembly technology of the lipid aptamer can realize cell 'pointing' gene modification and in-vivo gene reprogramming of cells.

Disclosed are CAR polypeptides comprising an antigen binding domain, a transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises a variant CD3 zeta (CD3ζ). Disclosed are nucleic acid sequences capable of encoding any of the disclosed CAR polypeptides. Disclosed are vectors comprising the nucleic acid sequence of the disclosed CAR nucleic acid sequences. Disclosed are cells comprising any of the CAR polypeptides, CAR nucleic acid sequences, or vectors disclosed herein. Disclosed are methods of treating a subject having cancer comprising administering a therapeutically effective amount of a composition comprising a T cell genetically modified to express one or more of the CAR polypeptides disclosed herein to the subject having cancer. Disclosed are methods of using one or more of the disclosed CAR polypeptides.

Provided herein are recombinant microorganisms that express a subject polypeptide. Microorganisms can comprise an expression construct comprising a flagellin promoter operatively linked with a heterologous nucleotide sequence encoding the subject polypeptide. The flagellin promoter sequence can comprise a genetic modification that reduces CsrA inhibition of translation. Microorganisms also can comprise a genetic modification that reduces FlgM inhibition of SigD initiation of transcription. The target polypeptide can be an aldehyde dehydrogenase. Such microorganisms are useful in the treatment of alcohol hangover.

The invention relates to a stem cell pretreatment technology in the field of biomedicine, which is characterized in that stem cells are pretreated in a specific parameter window (10-80mu g / mL, 5-60min and 36-38 DEG C) by establishing an ozone concentration-time-temperature cooperative control model, so that the anti-apoptosis ability, homing efficiency and paracrine function of the stem cells are remarkably enhanced. The method breaks through the technical bottleneck of low survival rate of traditional stem cell transplantation, and avoids the carcinogenic risk of gene modification. The application covers the three fields of myocardial repair, liver regeneration and nerve injury, and the special culture medium and the kit developed in a matched manner realize standardized operation. The survival rate of the pretreated stem cells after transplantation is increased by more than two times, the VEGF secretion amount is increased by 150%, and a safe and efficient new treatment strategy is provided for clinic.

The invention relates to the technical field of gene vectors, and discloses a multi-target multi-gene stable silencing vector and an application thereof in treating diabetes mellitus, and a multi-gene multi-target gene modification combined element is safely and efficiently integrated at a gene safety parking point of a second intron region of a CCR5 gene in a genome in a reverse fixed point manner by using a non-viral vector system; therefore, the effect of continuously silencing expression of a plurality of target genes such as MSTN, GRB10 and PTBP1 for a long time is achieved, and aging of mesenchymal stem cells cultured in vitro is remarkably inhibited. When the mesenchymal stem cells modified by the multi-target multi-gene stable silencing vector are infused, the activity is high, the chronic inflammatory microenvironment is effectively inhibited, and the senescence process is delayed; moreover, proliferation and differentiation of pancreatic beta cells are promoted, insulin secretion is enhanced, insulin resistance is effectively resisted, and diseases such as diabetes mellitus are prevented.

The invention concerns homogenous antibody-conjugates with high payload loading (high DAR) obtained by site-specific conjugation to a single antibody N-glycan. The conjugates according to the invention are homogeneous, i.e. have a DAR at or close to the theoretical DAR with a narrow distribution, and do not require any genetic modification of the antibody. The invention further concerns a modular, non-genetic preparation method for such conjugates, involving three simple steps and starting from any antibody. These steps are (a) enzymatic remodeling of the glycan to give an antibody functionalized with two or four click probes per antibody, (b) strain-promoted cycloaddition with a multivalent, bifunctional reagent comprising one cyclic alkyne and at least two click probes that are not reactive towards the cyclic alkyne, and (c) inverse electron-demand Diels-Alder cycloaddition of the click probes with branched linker-drug constructs comprising one cyclic alkyne or strained alkene, connected to one or more payloads preferably connected through a cleavable linker. The resulting conjugates, with DAR6 or higher, are rapidly generated with high homogeneity and with surprising stability. In addition, HIC profiles of the resulting ADCs indicate small relative retention time and therefore show high potential in the targeting of tumour cells and / or the treatment of cancer.

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.