73 results about "Gene Modification" patented technology

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.

Disclosed herein are genetically engineered Kluyveromyces marxianus cells capable of producing D-allulose with increased talitol formation. The engineered cell may comprise a genetic modification resulting in overexpression of a native talitol dehydrogenase enzyme; an exogenous polynucleotide sequence encoding an allulose-6-phosphate 3-epimerase enzyme at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identical to at least one of SEQ ID NOs:249-256, 258, and 259; and an exogenous polynucleotide sequence encoding an allulose-6-phosphate phosphatase enzyme at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identical to at least one of SEQ ID NOs:87, 89, 190, 123, 105, 107, 115, 83, 95, 113, 117, 119, 121, 127, 131, 137, 145, 169, 173, 179, and 183.

The process provided in this invention transfects primary cells with gene editing reagents using a high-volume gas-permeable cell culture device and a flow-through electroporation device under conditions that improve gene editing performance, cell yield, and drug (DP) quality characteristics for cell therapy applications. As demonstrated in the examples, primary cells edited according to the process provided herein achieved improved double-strand break (DSB) formation rates, increased frequency of homology-directed repair (HR) and non-homologous end joining (NHEJ) combinations, increased frequency of bi-allelic and mono-allelic HR events, improved cell viability, proliferative capacity, and cell fitness after gene editing, and reduced manufacturing time.

The invention provides genetically modified non-human animals that express chimeric human / non-human MHC I polypeptide and / or human or humanized β2 microglobulin polypeptide, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified animals and methods of making the same. Methods of using the genetically modified animals to study various aspects of human immune system are provided.

Provided herein are methods and compositions related to mice that express human or humanized Foot receptors (FcaR) from an FcaR locus positioned in the mouse leukocyte receptor complex (LRC). In certain embodiments, such mice are useful for in vivo testing of therapeutic agents comprising a human IgA Fc (e.g., the testing of the pharmacokinetic and / or pharmacodynamic properties of such therapeutic agents and dosing regimens). Also provided herein are methods of using such mice, cells from such mice, methods of making such mice, and ES cells comprising the same genetic modifications as such mice. Provided herein are methods and compositions related to mice that express human or humanized Foot receptors (FcaR) from an FcaR locus positioned in the mouse leukocyte receptor complex (LRC). In certain embodiments, such mice are useful for in vivo testing of therapeutic agents comprising a human IgA Fc (e.g., the testing of the pharmacokinetic and / or pharmacodynamic properties of such therapeutic agents and dosing regimens). Also provided herein are methods of using such mice, cells from such mice, methods of making such mice, and ES cells comprising the same genetic modifications as such mice.

Disclosed herein is a genetically engineered cells expressing a chimeric antigen receptor and methods and uses thereof.

This invention provides improved compositions and methods for mRNA display libraries for the rapid production of stable recombinant high-affinity binders. [Solution] A composition, method, and use of recombinant viruses and / or recombinant viral vectors encoding distinct antibodies or antibody fragments prepared from a highly diverse nucleic acid library are disclosed. Preferably, the recombinant virus is a genetically modified low immunogenic virus, for example, an E2b deletion adenovirus. The highly diverse nucleic acid library is (1)V H -CDR1 / 2 sublibrary, (2) multiple V H -CDR3 sublibrary and (3)V L It includes or is derived from sublibraries, each of which includes multiple members. Preferably, each member of the sublibrary includes at least one random cassette having multiple degenerate base positions.

This invention relates to a method for preparing genetically modified cells. Specifically, this invention provides a method for preparing genetically modified immune cells. This cell preparation method can rapidly prepare immune cells of high quality, thereby ensuring clinical efficacy.

This invention discloses a method for preparing NK cells, the NK cells obtained thereby, and their applications. The method includes: sorting NK cells from peripheral blood of an allogeneic donor; culturing them in a serum-free medium containing CD3 monoclonal antibody, CD16 antibody, and IL-2, without the need for a feeder layer; preferably, IL-21 is added for amplification on days 3-5 of culture, and the cells are cultured for 20-40 days. This method yields non-genetically modified NK cells, among which CD56... bright The NK cells comprise >90% of the cell line and highly express a variety of activating and functional molecules. These NK cells effectively clear Aβ oligomers and senescent cells, and alleviate neuroinflammation. In treatment, intracranial injection achieves equivalent or better efficacy with only about 1 / 10th the dose administered intravenously, significantly clearing Aβ deposits in the brains of AD model mice and improving pathology.

The application relates to the technical field of genetically modified yeasts, in particular to a Yarrowia lipolytica with high homologous recombination efficiency and / or low lipid degradation capacity and a construction method thereof. The engineered Yarrowia lipolytica comprises a copper ion inducible promoter and a gene related to homologous recombination; the copper ion inducible promoter is selected from any one or more of the following: a pCTR2 promoter, a pMT2 promoter and a pUAS16MT2 constitutive promoter; wherein the gene related to homologous recombination is selected from any one or more of the following: a ylKU70 gene, a ylKU80 gene, a ylRAD51 gene, a ylRAD52 gene, a scRAD52 gene and a scRAD59 gene. The homologous recombination efficiency of the engineered Yarrowia lipolytica is significantly improved. Furthermore, the POX5 gene, the POX2 gene and / or the POX3 gene are knocked out, so that the lipid degradation capacity of the engineered Yarrowia lipolytica is significantly reduced.

Provided herein are methods for the activation and expansion of genetically-modified Tcells, such as tumor infiltrating lymphocytes. In some cases, cells of the embodiments can be used for the therapeutic treatment of human diseases, such as cancer.

The present application relates to a kind of gRNA targeted to non-human animal OPG gene, preparation method of OPG gene knockout non-human animal model.The preparation method of OPG gene knockout non-human animal model includes the following steps: using gene editing technology, destroy non-human animal OPG gene, for the gRNA targeted to non-human animal OPG gene includes one or two shown in SEQ ID NO:1 And / or SEQ ID NO:2.The present application designs the gRNA specifically targeted to non-human animal OPG gene, using Cas9 protein, OPG gene is knocked out, causes frame shift mutation, to reach the purpose of gene knockout to non-human animal.The method of the present application to non-human animal OPG gene modification is simple and easy to operate, short cycle, using the method of OPG model for research OPG gene physiological mechanism and molecular mechanism provides foundation, while providing important tool for research osteoporosis pathogenesis and drug development.

This invention belongs to the field of genetic engineering technology, specifically disclosing a method for preparing and applying gene-modified homozygous cells of chicken progenitor cells (PGCs). Using a dual-fluorescent marker gene, two donor plasmids carrying green fluorescent protein (EGFP) and red fluorescent protein (mCherry), respectively, are transfected into PGCs. Cells simultaneously expressing red and green fluorescence are obtained using flow cytometry, thus forming homozygous cell lines. Then, the dual-fluorescent marker gene is deleted using the Cre-LoxP system combined with electroporation, and residual LoxP fragments are deleted using ssODN as a donor template, achieving traceless gene editing in recipient chicken embryos. This invention does not introduce any exogenous genes, increasing the safety of gene-modified chickens and is of great significance for eliminating the biosafety risks associated with gene-edited chickens.

The present application relates to a preparation method and application of photoreceptor precursor cells of human embryonic stem cell-derived retinal organoids, and belongs to the field of biotechnology and stem cell engineering. The present application constructs a lentiviral vector carrying an exogenous target gene and infects human embryonic stem cells to obtain a stable expression of engineered stem cell clone group by screening; the clone group is subjected to three-dimensional differentiation, and sequentially undergoes the formation of a blastoid, neural epithelial induction and retinal layering stage to obtain a retinal organoid; finally, the co-expression of photoreceptor precursor cell markers and exogenous genes in the organoid is identified. The present application realizes stable integration of genes at the source of stem cells, avoids the problem of low infection efficiency in the later stage, and provides a complete standardized scheme from gene modification, directional differentiation to function verification. The engineered photoreceptor precursor cells obtained by the method have important application value in the construction of retinal disease models, drug screening and cell therapy development.

The present disclosure provides compositions and methods for the genetic modification of cells, including, but not limited to, resting T-cells and hepatocytes. The compositions and methods can comprise lipid nanoparticles, wherein the lipid nanoparticles comprise at least one multivalent cationic bolaform amphiphilic lipid, at least one structural lipid, at least one phospholipid and at least one PEGylated lipid.

Compositions and methods for editing, e.g., altering, DNA sequences within TGFβR2 are provided. Compositions and methods for reducing or eliminating TGFβR2 protein expression in a cell are provided. Compositions and methods for immunotherapy are provided.

This invention relates to the field of biomedical technology, specifically disclosing a gene modification method for enhancing TCR-T cell activity and its application in tumor treatment. The method includes the following steps: T cells are activated in vitro; gene editing is performed to knock out endogenous immune checkpoint genes; a recombinant expression vector is constructed, the vector containing a polycistronic expression cassette driven by a single promoter, wherein the nucleic acid sequence encoding TCR and the nucleic acid sequence encoding an immunomodulatory factor are tandemly linked by a self-cleaving peptide coding sequence; the recombinant expression vector is introduced into the gene-knockout T cells; and the transduced T cells are cultured and expanded to obtain TCR-T cells with enhanced activity. This invention significantly enhances the anti-tumor activity, in vivo persistence, and tumor infiltration capacity of TCR-T cells through the synergistic effect of knocking out immune checkpoint genes and autocrine immunomodulatory factors; its modular design allows for flexible adaptation to different tumor antigen targets.

Disclosed are genetically modified rodents whose immunoglobulin heavy chain constant region loci are engineered such that a continuous segment between the CH1 exon and the CH3 exon of the endogenous IgH Gl is retained in the immunoglobulin heavy chain constant region loci of the rodents, but the CH1 exon of the endogenous IgH Gl is not expressed. Also provided are methods of making the genetically modified rodents and uses of the rodents.

The application discloses an sgRNA capable of effectively editing a pig CCR5 gene and application thereof, and is premised on an sgRNA capable of specifically recognizing a pig CCR5 gene in a pig genome, and successfully constructs a pig fetal fibroblast cell, a pig kidney cell line PK15 cell line and a pig 3D4 / 21 cell line with EGFP site-directed integration by using CRISPR / Cas9 mediated gene knock-in technology, and the results show that the EGFP gene can be stably and efficiently expressed in the three kinds of cells or cell lines. The application provides more site selection and reference for gene function research, gene modified pig research and development.

Provided herein are genetically modified cells such as pluripotent stem cells and mesenchymal stem cells and methods of making the same. The gene editing strategies of the present disclosure enable high expression of transgenes in genetically modified cells.

This invention belongs to the field of microbial fermentation technology, specifically a process for producing EPA using Schizochytrium fungi as raw material. It employs a dual-gene modified strain with MAT gene overexpression and CLF gene precise knockout, which is domesticated under high-sugar oxidative stress to obtain a highly stable strain. Through three-stage temperature-controlled, stepwise dissolved oxygen fermentation, and the combined addition of para-aminobenzoic acid and ascorbic acid, EPA synthesis is enhanced while oxidative loss is reduced. Highly efficient EPA enrichment is achieved through low-temperature ultrasonic cell disruption of wet cells, urea inclusion, two-stage molecular distillation, and tandem purification using immobilized composite adsorption decolorization. This invention significantly improves EPA yield and proportion, reduces oxidative loss, and enhances product purity and stability through the synergistic effect of gene modification, metabolic regulation, and purification processes. The process is highly controllable, suitable for industrial production, and yields EPA products with high purity and good stability, which can be directly used in the food, health product, and pharmaceutical fields.

This invention discloses a method for constructing an engineered algal strain of high-intensity sweet protein variants derived from *Chlamydomonas reinhardtii* and its application. First, the sweeteners Thaumatin and Brazzein are genetically modified. Then, through codon optimization based on *Chlamydomonas reinhardtii* preference, the encoding genes for CrThaumatin and CrBrazzein are synthesized and cloned into the expression vector pGM6. The recombinant expression vector is introduced into wild-type *Chlamydomonas reinhardtii* sp. algal strains. Using the paromomycin resistance marker carried by the expression vector, the engineered algal strains are successfully obtained. The engineered algal strains are fermented to produce dried, stable *Chlamydomonas reinhardtii* powder rich in the target sweet protein. Finally, oral administration tests after protein purification show that its sweetness is 3-5 times that of natural Thaumatin and Brazzein proteins.

This invention discloses a method for regulating cucumber seed germination at low temperatures. CsGAI Mutated genes and their applications. This invention constructs a targeted gene... CsGAI The gene editing vector pCas9CsGAI was used to transform cucumbers, and the resulting plasmid was then transformed into cucumbers. CsGAI Genetically modified plants were used for field hybridization and pollination, molecular identification, and phenotypic identification to prove that when CsGAI CsGAI CsGAI CsGAI CsGAI CsG When gene function is lost or weakened, the relative germination rate, relative germination potential, and relative germination index of cucumbers increase, and their cold tolerance improves. This invention lays the foundation for revealing the genetic and biological mechanisms of cold tolerance during cucumber germination, provides new gene resources for the cultivation of cold-tolerant cucumbers, and is of great significance for cucumber cold research and the breeding of new cold-tolerant varieties.

The present application provides a non-human animal expressing a human or chimeric sialic acid-binding Ig-like lectin 6 (SIGLEC6) protein and methods of using the same. In certain embodiments, the genome of the non-human animal comprises an exogenous polynucleotide sequence encoding a human or chimeric SIGLEC6 protein.

The present invention features genetically modified immune cells comprising novel adenosine base editors (e.g., ABE8) having enhanced anti-neoplasia activity, resistance to immune suppression, and decreased risk of eliciting a graft-versus-host reaction or host-versus-graft reaction, or a combination thereof. The present invention also features methods for producing and using these modified immune effector cells.

The invention provides compositions and methods for treating diseases associated with expression of EGFRvII. The invention also relates to chimeric antigen receptor (CAR) specific to EGFRvII, vectors encoding the same, and recombinant T cells comprising the anti-EGFRvIII CAR. The invention also includes methods of administering a genetically modified T cell expressing a CAR that comprises an anti-EGFRvIII binding domain.

This invention provides a non-human animal expressing a human or chimeric (e.g., humanized) NTCP protein and a method of using the same.

The present disclosure concerns recombinant yeast host cells having a first genetic modification for downregulating a first metabolic pathway that converts NADP+ to NADPH, as well as a second genetic modification for upregulating a second metabolic pathway that converts NADP+ to NADPH. The second genetic modification allows the expression of a glyceraldehyde-3-phosphate dehydrogenase lacking phosphorylating activity, which can, in some embodiments, be from enzyme commission 1.2.1.9 or 1.2.1.90. The second pathway is distinct from the first metabolic pathway. The present disclosure also concerns a process for making and improving the yield of a fermented product, such as ethanol, using the recombinant yeast host cell.

This disclosure relates to CRISPR-related systems and components for targeting, editing, and / or regulating the expression of the FLI-1 (Friend virus leukemia integration 1 transcription factor, Fli-1 proto-oncogene, ETS transcription factor) gene. This disclosure also relates to methods and applications relating to genetically modified cells, including T cells or T cell progenitor cells.