20 results about "Mammalian gene" patented technology

Disclosed herein are compositions of transcription activator-like effectors transcription factors and methods of using the compositions for inducing gene expression of mammalian genes.

The application discloses a codon-optimized Cas12i3 protein coding gene and application thereof. Specifically disclosed is a DNA molecule, and a nucleotide sequence of the DNA molecule is SEQ ID No. 3. The application also discloses a gene editing system containing the DNA molecule and a method for improving the efficiency of Cas12i3 protein-mediated gene editing. In order to better apply the Cas12i3 protein to mammalian gene editing, different codon optimization schemes are designed, and through software evaluation and experimental verification, a codon optimization scheme with good Cas12i3 expression and editing effect is compared and screened. The application screens the codon optimization scheme for efficiently expressing the Cas12i3 protein in mammals, which can greatly improve the editing efficiency of the Cas12i3 protein, lays a foundation for the wide application of the Cas12i3 protein in mammals, and has great practical application value.

Probe compositions and kits for identifying or assisting in the identification of mammalian species are provided. The probe composition is a nucleotide probe shown in SEQ ID NO: 1-SEQ ID NO: 3590 in the sequence listing. The COI gene of the target species is captured by the probe, and the DNA sequence of the COI gene is obtained by using the method of high-throughput detection of next-generation sequencing for library construction. It can take into account the advantages of flexibility and convenience of first-generation sequencing and can obtain results quickly, while solving the problem of cumbersome operation of batch samples in first-generation sequencing, and avoiding the risk of inaccurate COI gene sequences caused by unstable partial results in first-generation sequencing. Developing the probe into a mammalian COI gene capture kit can achieve rapid and batch identification of mammalian species.

The invention provides a retrotransposase, which comprises a target DNA binding structural domain containing a zinc finger binding motif, a reverse transcriptase structural domain and an endonuclease structural domain, and can be used for reversely transcribing RNA (Ribonucleic Acid) into DNA. The invention also relates to a system for modifying DNA. The system comprises the retrotransposase provided by the invention or a nucleic acid encoding the retrotransposase provided by the invention; and a donor RNA or a nucleic acid encoding the donor RNA, the donor RNA comprising: a sequence that binds to the retrotransposase and a heterologous sequence.

This application discloses a smart method for preparing gene-edited mammalian cells. The method includes: acquiring the target gene sequence information of the mammal to be observed and processing it according to preset screening rules to obtain the target sgRNA sequence; mixing an RNP complex suspension prepared based on the target sgRNA sequence with a suspension of embryonic stem cells to be edited, and performing electroporation transfection using multiple consecutive micropulses to obtain a transfected cell suspension; acquiring raw images of the transfected cell suspension and preprocessing them to obtain single-cell images, and using a trained lightweight convolutional neural network model to obtain the predicted gene-editing efficiency and prediction confidence after 48 hours; if the transfected cell suspension meets the quality control conditions, expanding and culturing the transfected cell suspension to obtain the prepared gene-edited cells. This method can shorten the cell culture cycle, reduce the waste of cell materials, improve the stability of transfection efficiency, and achieve precise preparation of gene-edited cells.

The invention discloses application of an SLC5A3 gene to improvement of swine AFB1 toxin tolerance, and belongs to the technical field of biology. It is found for the first time that SLC5A3 gene knockout can inhibit aflatoxin B1 from inducing apoptosis of mammal intestinal epithelial cells and improve the aflatoxin tolerance of the swine intestinal epithelial cells; as a result, the mammalian SLC5A3 gene is related to aflatoxin-induced intestinal injury, and may become a key target for breeding AFB1 pollution resistant livestock and poultry and a therapeutic target for toxin tolerance of mammalian intestinal diseases.

The present invention relates to a method, in particular an in vitro method, for identifying immune cells in a sample, comprising analyzing the methylation status of at least one CpG position in the mammalian gene region for GATA Binding Protein 3 (GATA3) according to SEQ ID No. 1, wherein a demethylation or lack of methylation of said at least one CpG position in the gene region is indicative for T helper cells, cytotoxic T cells, Th2 cells, memory CD4+ T cells or memory CD8+ T cells, in particular Th2 cells. The analysis according to the invention can identify immune cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood, non-blood or other immune cells. The present invention furthermore provides an improved method for quantifying immune cells, in particular in complex samples. The method can be performed without a step of purifying and / or enriching cells, preferably in whole blood and / or non-trypsinized tissue.

Embodiments of the disclosure concern the use of expression constructs in which at least one polyA signal is embedded upstream of an expressible transcript, such as within a 5′ UTR for the transcript, for example. In certain embodiments, the polyA signal is comprised within a ligand-binding aptamer, and the binding of the ligand to the aptamer, or lack thereof, dictates the outcome for the expressible transcript. In specific embodiments, absence of the ligand causes the expressed transcript having a polyA in its 5′ UTR to be expressed but then degraded, whereas presence of the ligand causes inhibition of degradation upon expression of the expressible transcript. More than one ligand-binding aptamer may be present on the same expression construct.

The present invention relates to a method, in particular an in vitro method, for identifying immune cells, comprising analyzing epigenetic modifications / properties of (including the methylation status) of at least one CpG position in the mammalian gene region for Integrin subunit alpha 4 (ITGA4), wherein a demethylation or lack of methylation of said at least one CpG position in the gene region to at least about 10% is indicative for ITGA4-positive cells, NK cells, and eosinophils, when compared to a CD4+ T cell, CD8+ T cell, monocyte, B cell, neutrophil, pDC, or ITGA4-negative cell. The analysis according to the invention can identify immune cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood, non-blood or other immune cells. The present invention furthermore provides an improved method for quantifying immune cells, in particular in complex samples. The method can be performed without a step of purifying and / or enriching cells, preferably in whole blood and / or non-trypsinized tissue.

Provided is a retrotransposase which comprises a target DNA binding domain comprising a zinc finger binding motif, a reverse transcriptase domain, and an endonuclease domain, and which is capable of reverse transcription of RNA to DNA. The invention also relates to a system for modifying DNA. The system comprises the retrotransposase or a nucleic acid encoding the retrotransposase; and a donor RNA or a nucleic acid encoding the donor RNA, the donor RNA comprising: a sequence that binds to the retrotransposase and a heterologous sequence.

The invention relates to the technical field of gene expression regulation, and discloses an iPsiR-based gene precise regulation expression system, which comprises: (a) a chimeric repressor protein, which is formed by fusing a PsiR protein and a repressor protein KRAB; (b) an artificial hybrid promoter, which comprises a human cytomegalovirus core promoter (PhCMV) and 3 copies of a PsiR operon sequence OPsiR connected in series at the downstream. By optimizing the artificial hybrid promoter, the expression of the PsiR gene in a eukaryotic system, especially in mammalian cells, is realized, and the successful breakthrough application of a psicose-induced gene regulation expression system in mammalian cells is promoted; therefore, the psicose-induced mammalian gene expression system has the application prospect of gene treatment of diseases.

This invention belongs to the field of biotechnology and discloses a high-yield rAAV plasmid system and its applications. The high-yield rAAV plasmid system of this invention includes a transgenic plasmid containing two terminal inverted repeat sequences, a packaging plasmid, a helper plasmid, and an shRNA expression cassette; the shRNA expression cassette includes a sequence capable of targeting the target gene in the transgenic plasmid. In the rAAV production process, the plasmid system of this invention inhibits the transcriptional expression of the target gene by expressing shRNA to target the coding region or target sequence of the target gene on the transgenic plasmid. The carefully designed target sequence, unlike coding regions of human and mammalian genes, can be broadly used in various transgenic plasmids, thereby increasing rAAV yield and significantly improving the rAAV production efficiency of mammalian cells, achieving the technical effects of improved rAAV druggability and reduced production costs.

Embodiments of the disclosure concern the use of expression constructs in which at least one polyA signal is embedded upstream of an expressible transcript, such as within a 5′ UTR for the transcript, for example. In certain embodiments, the polyA signal is comprised within a ligand-binding aptamer, and the binding of the ligand to the aptamer, or lack thereof, dictates the outcome for the expressible transcript. In specific embodiments, absence of the ligand causes the expressed transcript having a polyA in its 5′ UTR to be expressed but then degraded, whereas presence of the ligand causes inhibition of degradation upon expression of the expressible transcript. More than one ligand-binding aptamer may be present on the same expression construct.

The present disclosure provides, inter alia, codon-altered polynucleotides encoding Factor VIII variants for expression in mammalian cells.In some embodiments, the present invention also provides mammalian gene therapy vectors and methods for treating hemophilia A.

The present invention relates to a method, in particular an in vitro method, for specifically identifying the C-X-C motif chemokine receptor 3 (CXCR3) subpopulation of immune cells in a sample from a mammal comprising immune cells, in particular at least one of CD8+ effector and memory T cells, T helper (Th)1 cells and natural killer T (NKT) cells, comprising analyzing epigenetic modifications / properties of (including the methylation status) of at least one CpG position in the mammalian gene region for CXCR3 according to SEQ ID No. 1, wherein a demethylation or lack of methylation of said of at least one CpG position in said gene region is indicative for said CXCR3 specific subpopulation of immune cells in said sample, when compared to other immune cells, in particular for at least one of CD8+ effector and memory T cells, T helper (Th)1 cells and natural killer T (NKT) cells. The analyses according to the invention can identify the above cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The present invention furthermore provides an improved method for quantifying the above cells, in particular in complex samples. The method can be performed without a step of purifying and / or enriching cells, preferably in whole blood and / or non-trypsinized tissue.

The present disclosure provides, among other aspects, codon-altered polynucleotides encoding Factor VIII variants for expression in mammalian cells. In some embodiments, the disclosure also provides mammalian gene therapy vectors and methods for treating hemophilia A. In some embodiments, the present disclosure provides methods for dosing a hemophilia A patient with a polynucleotide, e.g., a codon-altered polynucleotide, encoding a Factor VII polypeptide.

The present invention relates to a method, in particular an in vitro method, for identifying memory B cells, comprising analyzing epigenetic modifications / properties of (including the methylation status) of at least one CpG position in the mammalian gene region for Chromobox protein homolog 6 (CBX6), wherein a demethylation or lack of methylation of said gene region is indicative for a memory B cell, when compared to a non-memory B cell. The analyses according to the invention can identify memory B cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The resent invention furthermore provides an improved method for quantifying memory B cells, in particular in complex samples. The method can be performed without a step of purifying and / or enriching cells, preferably in whole blood and / or non-trypsinized tissue.

The present invention relates to a method, in particular an in vitro method, for identifying immune cells, comprising analyzing epigenetic modifications / properties of (including the methylation status) at least one CpG position in the mammalian gene region for lymphocyte activating gene 3 (LAG3), wherein a demethylation or lack of methylation of said at least one CpG position in the gene region to at least about 50% is indicative for activated CD8+ T cells, memory CD8+ T cells, and LAG3-positive T cells, when compared to a granulocyte, monocyte, NK-cell, LAG-3 negative T cell, memory CD4+ T cell, naive CD4+ T cell or naive CD8+ T cell, or activated CD4+ T cell. The analysis according to the invention can identify immune cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood, non-blood or other immune cells. The present invention furthermore provides an improved method for quantifying immune cells, in particular in complex samples. The method can be performed without a step of purifying and / or enriching cells, preferably in whole blood and / or non-trypsinized tissue.

The present invention relates to a method, in particular an in vitro method, for identifying immune cells, comprising analyzing epigenetic modifications / properties of (including the methylation status) of at least one CpG position in the mammalian gene region for Prostaglandin D2 receptor 2 (PTGDR2), wherein a demethylation or lack of methylation of said at least one CpG position in the gene region to at least about 80% is indicative for eosinophils or basophils, when compared to a CRTH2- or CRTH+ leukocyte, CD8+ T cell, CD4+ T cell, CD14+ monocyte, CD19+ B cell, BDCA4+ pDC, CD56+ NK cell, and / or CD15+ granulocyte. The analysis according to the invention can identify immune cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood, non-blood or other immune cells. The present invention furthermore provides an improved method for quantifying immune cells, in particular in complex samples. The method can be performed without a step of purifying and / or enriching cells, preferably in whole blood and / or non-trypsinized tissue.

The present disclosure provides, among other aspects, codon-altered polynucleotides encoding Factor VIII variants for expression in mammalian cells. In some embodiments, the disclosure also provides mammalian gene therapy vectors and methods for treating hemophilia A.