62 results about "Genetically modified mouse" patented technology

Disclosed herein are non-human animals (e.g., rodents, e.g., mice or rats) genetically engineered to express a human or humanized T cell receptor (TCR) from a human or humanized TCR locus comprising a non-human TCR non-coding sequence, and optionally a humanized T cell co-receptor (e.g., humanized CD4 and / or CD8 (e.g., CD8α and / or CD8β)), and / or a human or humanized major histocompatibility complex that binds the humanized T cell co-receptor (e.g., human or humanized MHC II (e.g., MHC II α and / or MHC II β chains) and / or MHC I (e.g., MHC Iα) respectively, and optionally human or humanized β2 microglobulin). Also provided are embryos, tissues, and cells expressing the same. Methods for making the genetically engineered animals are also provided. Methods for using the genetically engineered animals for developing human therapeutics are also provided.

The present disclosure provides a method of evaluating a mechanism of bipolar disorder, in particular mood switch, in human-derived neurons which may include providing a mechanism of an attenuated protein interaction of CaV1.2 and BK that can cause the calcium signaling deficit, the main cellular phenotype. The present disclosure further provides a method of evaluating a composition for efficacy in treating bipolar disorder, including providing a genetically modified mouse exhibiting BK channel haploinsufficiency; administering the composition to the genetically modified non-human animal; exposing the genetically modified mouse animal to a mild stressor; and determining whether the genetically modified mouse animal experiences a behavioral switch. The present disclosure also provides a method of treating bipolar disorder in a subject in need thereof, including administering to the subject a therapeutically effective amount of a BK channel agonist.

The invention discloses construction of an alcoholic liver disease animal model of a GATA4 gene and application of the alcoholic liver disease animal model in prevention and treatment drugs, and belongs to the technical field of mouse model construction. The construction method of the animal model comprises the following steps: hybridizing a GATA flox transgenic hybrid mouse and a Vill-Cre transgenic mouse to obtain an F1 generation, selfing the transgenic mice with the genotypes of GATA4flox / + and Vill-Cre in the F1 generation to obtain an F2 generation mouse, identifying the genotypes of the mice by using a polymerase chain reaction (PCR) amplification reaction system, and screening to obtain a GATA4flox / flox / Vill-Cre transgenic hybrid mouse with the genotype of GATA4flox / + and Vill-Cre transgenic hybrid mouse with the genotype of GATA4flox / + and Vill-Cre transgenic hybrid mouse with the genotype of GATA4flox / + and Vill-Cre transgenic hybrid mouse. A GATA4 intestinal specific knockout mouse of the Vill-Cre is obtained; then, a Lieber-DeCarli liquid alcohol feed is given to the GATA4 intestinal specific knockout mouse, and after 7 weeks, the alcoholic liver injury mouse model with the GATA4 intestinal specific knockout is obtained. The invention provides a construction method of an alcoholic liver disease animal model with intestinal GATA4 gene specific knockout and application of GATA4 serving as a target spot in preparation of drugs for preventing and treating liver injury, liver diseases and other related diseases caused by alcohol, and the construction method has important clinical prospects.

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that present a choice of two human light chain variable gene segments such that the immunoglobulin light chains expresses by the mouse comprise one of the two human light chain variable gene segments. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided.

The present disclosure provides a genetically modified mouse comprising a genomic nucleic acid encoding human APOE4, a genomic nucleic acid encoding mouse TREM2 modified to include a R47H substitution, and at least one genomic modification selected from the group consisting of: (a) a genomic nucleic acid encoding mouse ABCA7 modified to include an A 1541 G substitution; (b) a genomic nucleic acid encoding mouse APP modified to include G60IR, F606Y, and R609H substitutions; (c) a genomic nucleic acid encoding mouse PLCG2 modified to include a M28L substitution; (d) a genomic nucleic acid encoding mouse MTHFR modified to include a A262V substitution; (e) an inactivated Ceacaml allele; and (f) an inactivated II1rap allele. Methods of producing the genetically modified mouse and methods of using the genetically modified mouse are also provided.

The invention discloses a construction method of a conditional knock-down dynactin animal model of astrocytes and an application of the animal model of the conditional knock-down dynactin animal model of the astrocytes. According to the method, a Dctn1LoxP gene knock-in mouse is hybridized with an Aldh1l1-Cre / ERT2 transgenic mouse, a target genotype mouse is obtained through three rounds of breeding, 100mg / kg Tamoxifen is continuously injected into the intraperitoneal cavity of the 2-month-old mouse for 5 days, and specific knock-down of dynactin in brain and spinal astrocytes is realized, including knockout of p150Glue and reduction of DCTN4, p50 and Arp1alpha protein levels. The model has the advantages of being high in specificity, permanent in intervention aging and capable of covering multiple life stages, the defects of a traditional model are overcome, the model can be used for researching the influence of dynactin on astrocyte neurobiological functions, a reliable tool is provided for screening related targets of nervous system diseases, and the model has important application value.

Genetically modified mice whose immunoglobulin loci are engineered to be inserted into gene segments of human immunoglobulin variable regions are disclosed, the mice being capable of normal reproduction and producing human-mouse chimeric antibodies comprising a human variable region and a mouse constant region. The invention also provides a method for preparing the genetically modified mouse and application of the mouse.

Disclosed is a genetically modified mouse having an immunoglobulin heavy chain locus engineered to insert a gene segment of a human immunoglobulin heavy chain variable region. The mouse is able to reproduce normally and produce a human-mouse chimeric antibody comprising a human heavy chain variable region and a mouse constant region. Also provided is a method for preparing the genetically modified mouse and use of the mouse.

Disclosed herein are non-human animals (e.g., rodents, e.g., mice or rats), the non-human animal is genetically engineered to express a humanized or human T cell receptor (TCR) comprising a variable domain encoded by (a) at least one human TCR variable region gamma gene segment and a (human) TCR gamma constant region gene sequence and / or (b) at least one human TCR variable region delta gene segment and a (human) TCR delta constant region gene sequence. Also provided are embryos, tissues, and cells expressing the receptors. Also provided are methods of making genetically engineered animals expressing the humanized or human gamma and / or delta TCRs. Also provided are methods of developing human therapeutics using the genetically engineered animals that produce a substantially humanized T cell immune response.

A genetically modified non-human animal lacking the expression of Klhcd7b is described. Methods and compositions for disrupting, deleting, and / or replacing the Klhcd7b coding sequence are described. Genetically modified mice useful as models of hearing loss or extremely severe deafness are also described. Loss of function cells, tissues, and embryos genetically modified to include Klhcd7b are also described.

Genetically modified non-human animals and methods and compositions for making and using them are provided, wherein the genetic modification comprises a deletion in an immunoglobulin constant region CH1 gene (optionally a deletion in a hinge region) of an IgG, IgA, IgD, and / or IgE, and wherein the mouse is capable of expressing a functional IgM. Genetically modified mice are described, including mice having a functional IgM gene and modified to have a deletion of a CH1 domain and a hinge region in a heavy chain constant domain that is not an IgM, e.g., in an IgG heavy chain constant domain. Genetically modified mice that make human variable / mouse constant chimeric heavy chain antibodies (antibodies that lack a light chain), fully mouse heavy chain antibodies, or fully human heavy chain antibodies are provided.

Disclosed in the present invention is a genetically modified mouse in which the immunoglobulin locus is engineered to be inserted with a gene segment of a human immunoglobulin variable region. The mouse is capable of reproducing normally and producing a human-mouse chimeric antibody, wherein the human-mouse chimeric antibody contains a human variable region and a mouse constant region. Further provided in the present invention is a method for preparing the genetically modified mouse and the use of the mouse.

The present invention relates to a HIF2α gene mutant, a vector, and their use in constructing a NAFLD mouse model. Genetically modified mice constructed using this mutant can be stably propagated, facilitating practical studies of the pathogenic mechanisms of the HIF2α gene in mice with non-alcoholic fatty liver disease (NAFLD). Given the limited availability of human NAFLD livers and the constraints of medical ethics, the mouse model provided in this application will become an important tool in NAFLD research, providing a stable, heritable research model for studies of pathogenic mechanisms, treatments, and drug screening.

Disclosed herein are non-human animals (e.g., rodents, e.g., mice or rats) genetically engineered to express a humanized T cell co-receptor (e.g., humanized CD4 and / or CD8 (e.g., CD8α and / or CD8β)), a human or humanized T cell receptor (TCR) comprising a variable domain encoded by at least one human TCR variable region gene segment and / or a human or humanized major histocompatibility complex that binds the humanized T cell co-receptor (e.g., human or humanized MHC II (e.g., MHC II α and / or MHC II β chains) and / or MHC I (e.g., MHC Iα) respectively, and optionally human or humanized β2 microglobulin). Also provided are embryos, tissues, and cells expressing the same. Methods for making a genetically engineered animal that expresses at least one humanized T cell co-receptor (e.g., humanized CD4 and / or CD8), at least one humanized MHC that associates with the humanized T cell co-receptor (e.g., humanized MHC II and / or MHC I, respectively) and / or the humanized TCR are also provided. Methods for using the genetically engineered animals that mount a substantially humanized T cell immune response for developing human therapeutics are also provided.

The invention discloses a construction method of an endothelial cell knockout Cdc42 gene aggravated bleomycin induced pulmonary fibrosis mouse model, which comprises the following steps of: mating a Cdc42 gene conditional knockout mouse with a mouse (Tie2-CreER) of which the endothelial cell specifically expresses tamoxifen induced Cre recombinase to obtain a double-transgenic mouse (Cdc42fl / fl-Tie2-CreER); the Cdc42 gene is specifically knocked out in vascular endothelial cells under the induction of tamoxifen, and then pulmonary fibrosis is induced by subcutaneous injection of bleomycin. The model shows aggravated pulmonary fibrosis phenotypes, including collagen deposition increase, alveolar structure damage and fibrosis-related protein expression up-regulation, and is high in construction success rate and good in repeatability. The invention also provides an application of the model in research of systemic sclerosis related interstitial lung disease pathogenesis and screening of anti-pulmonary fibrosis drugs, and an application of the Cdc42 gene as a drug target, and provides an accurate and reliable tool for pulmonary fibrosis research.

A genetically modified mouse is provided, wherein the mouse is incapable of rearranging and expressing an endogenous mouse immunoglobulin light chain variable sequence, wherein the mouse expresses only one or two human light chain variable domains encoded by human immunoglobulin sequences operably linked to the mouse kappa (κ) constant gene at the endogenous mouse κ locus, wherein the mouse expresses a reverse chimeric antibody having a light chain variable domain derived from one of only two human light chain variable region gene segments and a mouse κ constant domain, and a human heavy chain variable domain and a mouse heavy chain constant domain, from an endogenous mouse heavy chain locus. Bispecific epitope-binding proteins that are fully human are provided, comprising two different heavy chains that associate with an identical light chain that comprises a variable domain derived from one of two different human light chain variable region gene segments.

Chimeric transgenic immunoglobulin (Ig) mice comprising altered heavy chain loci are provided in which endogenous mouse D and J segments have been deleted and at which a human heavy chain Ig transgene is inserted, such that the transgenic Ig mice express an antibody library that utilizes human VH and mouse VH in the heavy chain, each linked to human DH and JH segments, thus, enhanced diversity is produced. Methods of making and using transgenic animals (e.g., to produce antibodies) are also provided.

The invention provides a preparation method of a genetically modified mouse capable of mediating Tbx1 inherent disorder protein structural domain deletion. The method comprises the following steps: (a) constructing a targeting vector; (b) preparing a gene editing compound; (c) microinjection; (d) embryo transplantation and screening; (e) breeding and establishing a stable strain. Sequencing verifies that the gene modified mouse can stably express the Tbx1 mutant protein which lacks IDR and carries an N-terminal 3xFlag tag, and a homozygote mutant mouse shows remarkable developmental defects such as ventricular septal defect and craniofacial deformity; and an important genetic tool is provided for researching the action mechanism of the phase separation characteristic of the Tbx1 protein in the heart, craniofacial development and congenital heart disease (such as DiGeorge syndrome).

A mouse with a humanization of the mIL-3 gene and the mGM-CSF gene, a knockout of a mRAG gene, and a knockout of a mIl2rg subunit gene; and optionally a humanization of the TPO gene is described. A RAG / Il2rg KO / hTPO knock-in mouse is described. A mouse engrafted with human hematopoietic stem cells (HSCs) that maintains a human immune cell (HIC) population derived from the HSCs and that is infectable by a human pathogen, e.g., S. typhi or M. tuberculosis is described. A mouse that models a human pathogen infection that is poorly modeled in mice is described, e.g., a mouse that models a human mycobacterial infection, wherein the mouse develops one or more granulomas comprising human immune cells. A mouse that comprises a human hematopoietic malignancy that originates from an early human hematopoietic cells is described, e.g., a myeloid leukemia or a myeloproliferative neoplasia.

Disclosed in the present disclosure is genetically modified mice in which immunoglobulin loci are modified to insert gene segments of a human immunoglobulin variable region. The mice can be bred normally and produce human-mouse chimeric antibodies including a human variable region and a mouse constant region. The present disclosure also provides a method for preparing the genetically modified mice and use of the mice.

Disclosed herein is a genetically modified mouse whose genome comprises a transgene encoding a Miltenberger blood group antigen subtype III (Mi.II antigen). According to embodiments of the present disclosure, the Mi.III antigen comprises the amino acid sequence of SEQ ID NO: 1. Also disclosed herein are a method of producing the genetically modified mouse, and uses of the genetically modified mouse in selecting a drug candidate for treating hypertension.

The invention discloses a pancreatic cancer organoid with tumor microenvironment characteristics and a construction method and application thereof, and belongs to the technical field of organoid. According to the invention, KPC transgenic mouse in-situ pancreatic tumor tissue is adopted as a cell source, and an organ with a pancreatic cancer tissue structure, biological characteristics and a complex tumor microenvironment is successfully constructed in combination with a culture medium formula with optimized culture conditions and characteristics and a multi-component co-culture mode; the obtained organoid is composed of multiple cell types such as tumor cells, cancer-related fibroblasts (CAFs), tumor-related macrophages (TAMs), myeloid-derived suppressor cells (MDSCs), T cells and the like, and a complex pancreatic cancer immune microenvironment can be rebuilt truly. The organ can be widely applied to efficacy evaluation of pancreatic cancer immunotherapy drugs, an efficient and repeatable in-vitro experimental platform is provided for early-stage research and drug development of pancreatic cancer immunotherapy, the research and development process of related drugs is expected to be accelerated, and the clinical conversion efficiency is improved.

Bag3 is a multifunctional protein expressed predominantly in the heart, the skeletal muscle, the central nervous system and in many cancers. Although BAG3 was cloned only a decade ago, studies have shown that genetic variants, particularly those that result in haplo-insufficiency, can lead to severe left ventricular dysfunction; however, the full mechanisms responsible have remained obscure. To obviate the influence of heart failure itself on the biology of Bag3, transgenic mice harboring a single allele knock-out were studied between 8 and 10 weeks of age before any obvious signs of heart failure were evident. The results were surprising and informative. First, it was found that despite a normal phenotype, young Bag3+ / − had marked changes in the proteome that were characterized by changes in proteins associated with metabolism and apoptosis. Consistent with this finding, a decrease in the levels of critical proteins charged with maintaining the mitochondrial membrane potential was observed. It was also found that young mice shifted from a balance between the extrinsic and intrinsic pathways of apoptosis. However, in the presence of stress and the absence of Bag3 there was a shift from a balanced to an extrinsic dominant system (cleaved caspase 8). The diverse array of critical pathways regulated by Bag3 suggests a more important role especially during stress and that this role might include serving as an intracellular glue that holds proteins where they can be most effective rather than having them meet accidentally.

Disclosed in the present invention is a genetically modified mouse in which the immunoglobulin locus is engineered to be inserted with a gene segment of a human immunoglobulin variable region. The mouse is capable of reproducing normally and producing a human-mouse chimeric antibody, wherein the human-mouse chimeric antibody contains a human variable region and a mouse constant region. Further provided in the present invention is a method for preparing the genetically modified mouse and the use of the mouse.

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that present a choice of two human light chain variable gene segments such that the immunoglobulin light chains expresses by the mouse comprise one of the two human light chain variable gene segments. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided.

To provide a hybrid light chain mouse.SOLUTION: There is provided a mouse genetically modified to appear a Homo sapiens λ variable (hVλ) sequence (containing a mouse for appearing the hVλ sequence from an endogenous mouse λ light chain gene locus, a mouse for appearing an hVλ sequence from an endogenous mouse κ light chain gene locus, and a mouse for appearing hVλ sequence from a transducing gene or an episome). Here, the hVλ sequence is connected to a mouse steady sequence. There is provided a mouse for becoming an origin of the Homo sapiens λ variable sequence which has been varied into a body cell variation useful for preparing an antigen binding protein. There are provided a composition and a method for preparing an antigen binding protein containing a human λ variable sequence containing a human antibody.SELECTED DRAWING: Figure 7B

The present invention provides genetically modified animals (e.g., mice), humanized heavy chain antibodies, humanized nanobodies, and methods for producing and using them. [Solution] Provided are genetically modified non-human animals (e.g., genetically modified mice) that can be designed to produce heavy chain antibodies that can be used to generate single-domain antibodies or nanobodies. In one embodiment, a genetically modified mouse is provided comprising a germline modification comprising deletion of nucleic acid sequences comprising one or more heavy chain C region genes; the genetically modified mouse expresses an IgG heavy chain antibody and secretes an IgG heavy chain antibody in its serum.

The application discloses a construction method and application of an NLRP3 humanized mouse model, and belongs to the technical field of genetic engineering. The construction method of the NLRP3 humanized mouse model comprises the step of mutating the 708th amino acid of mouse Nlrp3-201 protein into a non-acidic amino acid; the Ensembl ID of the mouse Nlrp3-201 is ENSMUST00000079476.10. It is verified that the 708th amino acid of the mouse NLRP3 protein is mutated into a non-acidic amino acid (for example, alanine), the NLRP3 of the obtained mutant transgenic mouse is similar to human NLRP3, and cannot be cut by caspase-3, and humanization is realized. The mutant mouse model can better reflect the biological processes such as human inflammasome activation, cell pyroptosis and inflammatory reaction, and provides a more accurate animal model for understanding human diseases and drug screening.

Disclosed in the present disclosure is genetically modified mice in which immunoglobulin loci are modified to insert gene segments of a human immunoglobulin variable region. The mice can be bred normally and produce human-mouse chimeric antibodies including a human variable region and a mouse constant region. The present disclosure also provides a method for preparing the genetically modified mice and use of the mice.