13 results about "Chain gene" patented technology

The present invention discloses a genetically modified rodent whose immunoglobulin light chain locus is modified so as to comprise a single rearranged human immunoglobulin light chain gene V / J gene upstream of a mouse Kappa light chain locus constant region, a mouse variable region sequence cannot bind to an endogenous constant region to form a complete mouse light chain antibody, the rodents are capable of normally reproducing and producing human light chain-containing antibodies. The invention also provides a method of making the genetically modified rodent and a use of the rodent.

This invention provides an organ fibrosis-related micropeptide, OFMP, and its applications. This type of micropeptide, OFMP, is a translational product with endogenous biological activity and stable existence, encoded by an open reading frame (ORF) in a long non-coding RNA. It is the first time that the long non-coding RNA encoding the micropeptide has been found to be significantly related to organ fibrosis and can encode the micropeptide. Synthetic peptides prepared based on the aforementioned OFMP micropeptide show significant inhibitory effects on several key indicators of organ fibrosis, including type I collagen α1 chain gene, α-smooth muscle actin, transforming growth factor β1, and connective tissue growth factor. This indicates that the OFMP micropeptide, the nucleotide sequence (ORF) encoding the micropeptide, and the long non-coding RNA sequence containing the nucleotide sequence have important clinical diagnostic and therapeutic value for organ fibrosis.

Provided herein is a transgenic chicken comprising a genome having an endogenous immunoglobulin heavy chain gene in which the IgY CH1 coding sequence is functionally deleted. In certain embodiments, in B cells of the chicken the endogenous immunoglobulin heavy chain gene comprises: (a) a functional immunoglobulin heavy chain gene comprising, in operable linkage, a nucleic acid encoding an antibody heavy chain variable domain and a constant region coding sequence in which the IgY CH1 coding sequence is functionally deleted; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the variable domain of (a), wherein the pseudogenes are upstream of the nucleic acid encoding an antibody heavy chain variable domain of (a).

This disclosure provides, among other things, a transgenic animal that uses gene conversion for antibody diversification, comprising B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding an autonomous heavy chain (AHC) variable domain; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the AHC variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.

A composition for increasing the efficiency of genome editing with a single-strand-form editing polynucleotide capable of modifying a target site in a double-stranded genome DNA. The composition includes at least one protein selected from Hfq, DnaK, and variants thereof, or a nucleic acid having a base sequence that encodes the protein. A genome editing kit including the composition and an editing polynucleotide. A method for modifying a target site in a double-stranded genome DNA of a cell or a living organism including treating a cell or living organism with the genome editing kit. A method for producing a cell or living organism in which a target site in a double-stranded genome DNA has been modified.

Genetically modified non-human animals are provided that express an immunoglobulin variable domain that comprises at least one histidine, wherein the at least one histidine is encoded by a substitution of a non-histidine codon in the germline of the animal with a histidine codon, or the insertion of a histidine codon in a germline immunoglobulin nucleic acid sequence. Immunoglobulin genes comprising histidines in one or more CDRs, in an N-terminal region, and / or in a loop 4 region are also provided. Immunoglobulin variable domains comprising one or more histidines (e.g., histidine clusters) substituted for non-antigen-binding non-histidine residues. Non-human animals that are progeny of animals comprising modified heavy chain variable loci (V, D, J segments), modified light chain variable loci (V, J segments), and rearranged germline light chain genes (VJ sequences) are also provided. Non-human animals that make immunoglobulin domains that bind antigens in a pH-sensitive manner are provided.

The invention provides a preparation method of an ictalurus punctatus IgM antibody based on a prokaryotic expression system, and relates to the field of aquatic animal immunodetection and antibody preparation, and the preparation method comprises the following steps: designing a specific primer based on an ictalurus punctatus IgM protein gene sequence in GenBank, and carrying out antigen determinant truncation; the method comprises the following steps: extracting liver RNA (Ribonucleic Acid) of channel catfish by a Trizol method, reversely transcribing the liver RNA into cDNA (Complementary Deoxyribonucleic Acid), and cloning an IgM The method comprises the following steps: constructing a pET-30a-IgM recombinant expression vector, transforming Escherichia coli Rosetta competent cells, and inducing protein expression; purifying the truncated IgM protein by adopting the steps of ultrasonication, affinity chromatography and dialysis concentration; a New Zealand white rabbit is immunized to prepare antiserum, and the titer is detected through ELISA. According to the method, efficient and low-cost preparation of the IgM protein is realized by utilizing a prokaryotic expression system, the period is only several weeks, high-purity protein of milligram-grade to gram-grade can be obtained, the batch stability is good, related biological safety risks of animal serum are avoided, and a key basis is provided for development of an ictalurus punctatus ELISA (enzyme-linked immunosorbent assay) kit, immunohistochemistry and B lymphocyte research.

Adeno-associated virus (AAV) expression vectors for the treatment of Charcot-Marie-Tooth disease are provided. The AAV expression vectors can have at least one neurofilament light chain gene (NEFL)-targeting shRNA operably linked to a first promoter and a NEFL cDNA operably linked to a second promoter. Pharmaceutical compositions comprising AAV expression vectors are also provided, as are methods of treating Charcot-Marie-Tooth disease type 2E.

The present invention provides a method for producing a circular vector that includes a long-chain gene cluster without mutations, the method involving ligating, in one pod and in one step in the presence of an exonuclease, a DNA polymerase, a DNA ligase, and a master mix, a linear vector and three or more DNA fragments having an overlap of 40-60 bp on both ends and constituting a long-chain gene cluster.

Disclosed is a genetically modified rodent an immunoglobulin light chain locus of which is engineered such that a single rearranged human immunoglobulin light chain gene V / J gene is contained upstream of a constant region of the mouse Kappa light chain locus, and thus a mouse variable region sequence cannot bind to the endogenous constant region to form a complete mouse light chain antibody. The rodent can reproduce normally and produce an antibody containing a human light chain. Also provided are a preparation method for the genetically modified rodent and the use thereof.

To provide a method for simultaneously measuring amounts of TREC, KREC and SMN1, an inspection method using the same, and a kit thereof.SOLUTION: A method for simultaneously measuring amounts of T-cell receptor recombination excision circle (TREC), Igκ-deleting recombination excision circle (KREC) and SMN1 gene by quantitative PCR includes: (i) a process of mixing DNA and PCR reagent collected from a subject, where the PCR reagent includes a set of primers and a set of probes; and (ii) a process of performing quantitative PCR reaction to quantitatively determine the amounts of TREC, KREC and SMN1 gene, where the set of primers includes six primers including base sequences shown by sequence numbers 4, 5, 7, 10, 37 and 38, and the set of probes includes three probes including base sequences shown by sequence numbers 22, 23 and 28.SELECTED DRAWING: None

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.