19 results about "Totipotent" patented technology

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention relates to the technical field of biomedicine, in particular to a method for regulating and controlling the liquid-liquid phase separation capacity and function of DUX4 and application of the method, and the liquid-liquid phase separation capacity and function of the DUX4 are regulated and controlled by mutating an internal disordered region of the DUX4. The invention finds that the mutation has a remarkable inhibition effect on expression of DUX4 activated totipotent genes and FSHD related genes; according to the method, an intracellular liquid-liquid phase separation model is constructed through intracellular overexpression of DUX4; a research basis and a model are provided for subsequent LLPS research of DUX4, and thinking and mechanism explanation are provided for research of DUX4-mediated facial shoulder brachial muscular dystrophy (FSHD) and development of treatment targets.

The invention discloses a vector system and method capable of converting mouse embryonic stem cells into embryonic-like cells in a 2-cell period, the vector system comprises a mouse Zfp352 gene sequence, and the nucleotide sequence of the mouse Zfp352 gene is shown as SEQ ID No.1. The invention also discloses a method for converting the mouse embryonic stem cells into embryonic-like cells in a 2-cell period. The invention creatively discovers that through overexpression of the Zfp352 in the mouse embryonic stem cells, the efficiency of converting the mouse pluripotent embryonic stem cells into the totipotent embryonic-like cells in the 2-cell period can be remarkably improved.

The present invention relates to methods of determining a potency state of an embryonic stem cell (ESC) in culture, comprising detecting the expression level of Negative Elongation Factor Complex Member A (Nelfa) in the ESC, and correlating the expression level of Nelfa with that of a reference sample to determine the potency state of the ESC. The present invention also provides methods of selecting totipotent-like ESCs from a population of ESCs in culture using an anti-Nelfa antigen binding protein, methods of inducing totipotency in an ESC in culture, methods of improving the reprogramming efficiency of a somatic cell into an induced pluripotent stem cell and methods of reprogramming a somatic cell into a totipotent stem cell by inducing Nelfa expression. Cells comprising an expression vector encoding Nelfa and kits for use in the methods of the invention are also provided.

The invention discloses a PPR (pentatricopeptide repeats) high-salt-resistant all-potent nuclease mutant and application thereof. The invention firstly discloses an amino acid sequence of the PPR high-salt-resistant all-potent nuclease mutant as shown in SEQ ID NO. 1. The invention further discloses application of the PPR high-salt-resistant all-potent nuclease mutant to removal of nucleic acid. According to the PPR high-salt-resistant totipotent nuclease mutant, alanine at the 165 site of wild PPR high-salt-resistant totipotent nuclease which does not contain signal peptide is replaced with aspartic acid, overexpression can be carried out in a prokaryotic host, and a large number of proteins can be obtained through purification. The PPR high-salt-resistant all-purpose nuclease mutant disclosed by the invention maintains high activity in a wide salt concentration range, a temperature range, a pH range and a Mg < 2 + > range, and can be compatible with most additives used in a biological process. In addition, the activity of the PPR high-salt-resistant all-purpose nuclease mutant is superior to that of other domestic competitive products, and the competitive product level with the best commercial effect at present can be achieved.

The invention relates to the technical field of enzyme engineering, and particularly provides an all-round nuclease variant, the variant can tolerate a wider salt concentration range compared with wild type all-round nuclease, good enzyme activity is still kept under the increased salt concentration, and the application of all-round nuclease under different salt concentrations is widened.

The invention provides a compound capable of inducing totipotent embryonic stem cells, the structural general formula of the compound is shown in the specification, and the preparation method of the compound comprises the following steps: (1) preparing an intermediate Int-1, (2) preparing a Boc-removed product Int-2, and (3) preparing a product MYF. The micromolecule compound (MYF) can simultaneously activate human and mouse totipotent embryonic stem cells in vitro, and fills the technical blank that a single micromolecule can only activate human or mouse totipotent embryonic stem cells independently in the prior art.

The present disclosure relates to methods for generating progenitors functionally totipotent blastomere-like stem cells via P ARP -regulated proteomic reprogramming of a PARP- DUX4-NAN0G regulatory axis of conventional human induced pluripotent stem cells (hiPSC) by chemically reverting them to tankyrase / PARP1 inhibitor-regulated naive (TIRN) human stem cells and methods and compositions useful for generating chimeras using such human induced totipotent TIRN stem cells. In certain embodiments, the TIRN stem cells are engineered to express a cell adhesion molecule (e.g., E-Cadherin) or DUX4.

The invention provides a culture method for promoting conversion of mouse embryonic stem cells into totipotent stem cells. The cells can be kept in a brand new form and subcultured under the condition. According to the method adopted by the invention, an inhibitor WM-3825 of 2 mu M H4K16ac is added into a DMEM (Dulbecco Modified Eagle Medium) with a culture medium composition of 10% fetal calf serum according to a proportion of 1: 1000, the mouse embryonic stem cells can be normally cultured and passaged, the change of the totipotent gene is detected through real-time fluorescent quantitative PCR (Polymerase Chain Reaction), the expression of the totipotent related gene Dux is found to be obviously increased, and the expression of the mouse embryonic stem cells is obviously improved. Flow cytometry detection shows that the proportion of the totipotent stem cells is remarkably increased, and AP dyeing shows that the pluripotency of the mouse embryonic stem cells is reduced. The method starts from cell culture, and the method is possibly suitable for research on conversion from pluripotency to totipotency of embryonic stem cells of other mammals including human. By adding a small-molecule inhibitor WM-3825 of H4K16ac, a brand-new method for promoting conversion from the mouse pluripotent stem cells to the totipotent stem cells is established, and the 2-cell period of mouse embryonic development can be simulated in vitro through the culture system. The invention provides a new method for the generation of the totipotent stem cells, deepens the understanding of people on different states of the stem cells, and provides a new perspective for understanding an epigenetics mechanism between the conversion of the totipotent stem cells and the conversion of the pluripotent stem cells.

The present application provides a method for preparing human ovarian somatic cell-like cells. The present application establishes a method for preparing human ovarian somatic cell-like cells in vitro, which comprises stage-based culture based on human stem cells (including human pluripotent stem cells and human totipotent stem cells). The method of the present application can efficiently obtain functional human ovarian somatic cell-like cells, promote the differentiation ability of female germ stem cells (FGSC) cultured in vitro, and provide a new approach for assisted reproduction and treatment of infertility.

Provided herein is a composition comprising genetically modified totipotent / pluripotent stem cells. A method of xenotransplantation, and a system and a method of evaluating the suitability of a composition comprising totipotent / pluripotent stem cells for xenotransplantation are also provided.

The present invention relates to a method for producing hematopoietic stem cells derived from totipotent stem cells, and a method for producing a humanized mouse model using the produced hematopoietic stem cells. According to one aspect, the method for producing hematopoietic stem cells can highly efficiently differentiate hematopoietic stem cells from totipotent stem cells without gene insertion, and optimal differentiation conditions have been confirmed by combining low molecular weight compounds and protein growth factors.

The invention provides a method for promoting induction of human totipotent stem cells, belongs to the technical field of biology, and particularly relates to a method for promoting induction of the human totipotent stem cells to the totipotent stem cells through OTX2 knockout or knockdown, the method comprises the following steps: S1, constructing an OTX2 knockout or knockout plasmid vector; s2, establishing a human pluripotent stem cell line with OTX2 knocked down or knocked out; and S3, inducing the human pluripotent stem cells of which the OTX2 is knocked down or knocked out into totipotent stem cells, which are also called 8-cell-like cells (8CLCs). According to the invention, the efficiency of inducing the human pluripotent stem cells into the totipotent stem cells can be improved, and the application of the totipotent stem cells in in-vitro blastocyst-like induction, in-vivo chimeric embryo formation and the like is provided.

The invention relates to the use of a laminin (LN) matrix for hepatic differentiation. The invention also relates to a method for inducing hepatic differentiation comprising the steps of: (i) providing a population of human totipotent cells, (ii) culturing the population on a laminin-coated support in an endoderm stimulation medium to produce a population of human DE cells, (iii) culturing said population of human DE cells on a laminin-coated support in a hepatic stimulation medium to produce a population of human hepatoblast-like cells, and (iv) optionally culturing said population of human hepatoblast-like cells on a laminin-coated support in a hepatic maturation medium to produce a population of human hepatocyte-like cells.The invention further relates to a population of human hepatoblast-type cells or human embryonic hepatocyte-type cells obtained by the method of the invention. The invention further relates to a population of human hepatoblast-type cells expressing HNF4 α and expressing substantially AFP for use in a method of treating the human body.

The present invention relates to methods of determining a potency state of an embryonic stem cell (ESC) in culture, comprising detecting the expression level of Negative Elongation Factor Complex Member A (Nelfa) in the ESC, and correlating the expression level of Nelfa with that of a reference sample to determine the potency state of the ESC. The present invention also provides methods of selecting totipotent-like ESCs from a population of ESCs in culture using an anti-Nelfa antigen binding protein, methods of inducing totipotency in an ESC in culture, methods of improving the reprogramming efficiency of a somatic cell into an induced pluripotent stem cell and methods of reprogramming a somatic cell into a totipotent stem cell by inducing Nelfa expression. Cells comprising an expression vector encoding Nelfa and kits for use in the methods of the invention are also provided.

This invention discloses a method for inducing mouse embryonic stem cells to transition to a pluripotent state using H2BC21 and its application, belonging to the fields of molecular biology and stem cell engineering technology. By constructing an interference vector targeting the H2BC21 gene, packaging it with lentivirus, and infecting mouse embryonic stem cells, the expression of the H2BC21 gene is knocked down, and embryonic stem cell lines with stable low expression of H2BC21 are obtained through screening. This invention is the first to discover that H2BC21 is a core negative regulator of mammalian embryonic stem cell pluripotency. After knocking down H2BC21, the expression of pluripotency core genes Nanog, Oct4, and Sox2 is downregulated, while the expression of pluripotency core genes Zscan4b, Usp17la, and Tmem92 is upregulated. The cells exhibit molecular characteristics similar to 2-cell phase cells, successfully achieving the transition from pluripotency to a pluripotent state.

This invention discloses a method for preparing inclusion bodies based on dual-promoter expression plasmids and its application, belonging to the field of biotechnology. The invention includes screening of totipotent nucleases, purification of totipotent nucleases, verification of the degradation activity of the totipotent nuclease DNS on nucleic acids, construction of dual-promoter recombinant strains, fermenter culture, cell recovery, and cell disruption to obtain crude inclusion bodies. This invention achieves high-efficiency expression of the recombinant human growth hormone gene induced by the T7 promoter and high-activity expression of the totipotent nuclease gene DNS induced by the cspA promoter, demonstrating significant effects in the application of recombinant human growth hormone. During the preparation process, the cell viscosity is low, the homogenization and disruption effect is improved, the BL21(DE3) / DGH cell disruption rate is high, the inclusion body content harvested by centrifugation per unit volume of fermentation broth is high, and the residual amount of HCD (host DNA residue) in the target protein per unit mass is reduced, effectively reducing energy consumption.

The invention discloses construction and application of a vector for expressing totipotent nuclease and a host cell, and discloses a fusion signal peptide of which the nucleotide sequence is shown as SEQ ID NO.1. The transformed signal peptide pelb guides totipotent nuclease to be converted into periplasmic space of escherichia coli Rosetta (DE3) host cells for efficient secretory expression, and a matched purification process is suitable for scenes of vaccine production, host nucleic acid residue removal in biological pharmacy and the like.

This process yields totipotent plant cells capable of regenerating plantlets, similar to propagation, in vitro micropropagation, and synthetic seed production. It leverages spray drying as a technology for preserving plant germplasm and as a novel method not currently available. The process begins with somatic cells in suspension and involves inducing friable embryogenic calluses from seeds, leaves, stems, roots, or any plant explant capable of generating cells in suspension, followed by obtaining the cell pack. This cell pack is mixed with encapsulating agents and then spray-dried. Finally, the spray-dried cells are rehydrated and reactivated to continue their morphogenic development, resulting in plantlets capable of acclimatizing to greenhouse and field environments.