9 results about "Mouse Embryonic Stem Cell" patented technology

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 application provides a non-coding RNA combination for inducing totipotency of mouse embryonic stem cells and application thereof, and relates to the technical field of biology.The combination comprises: an enhancer RNA with a sequence shown as SEQ ID NO:1 or SEQ ID NO:2, and a long-chain non-coding RNA with a sequence shown as SEQ ID NO:3.The application discloses, for the first time, that the CRISPR activation system is used to synergistically up-regulate the expression of the combination, and the conversion of mouse embryonic stem cells to two-cell-like cells can be efficiently promoted.The application does not need to integrate exogenous transcription factors, and thus the risk of genome is avoided; the sgRNA is precisely targeted, and thus the defect of non-specific off-target of chemical small molecules is overcome; a significant synergistic effect is generated, the induction efficiency of two-cell-like cells is significantly improved to about 11.6%, and a new strategy which is safer, more efficient and controllable is provided for stem cell fate reprogramming and early development research.

The application provides a non-coding RNA combination and application thereof in mouse totipotent stem cell induction, and relates to the technical field of biotechnology.The combination comprises: an enhancer RNA with a sequence as shown in SEQ ID NO:1 and SEQ ID NO:2, and a long-chain non-coding RNA with a sequence as shown in SEQ ID NO:3.The application discloses, for the first time, that the expression of the endogenous non-coding RNA combination is synergistically inhibited by using a CRISPR inhibition system, and the mouse embryonic stem cells can be efficiently promoted to transform into 2C-like cells.The application does not need to integrate an exogenous transcription factor, avoids the safety risk of the genome, overcomes the defect of non-specific off-target of a chemical small molecule by precise targeting, and significantly improves the induction efficiency of the 2C-like cells to about 9.09%.The application provides a safer, more efficient and controllable new strategy for stem cell fate reprogramming and early development research.

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 invention relates to the technical field of cell culture, in particular to a method for induced culture of embryonic stem cells. The method comprises the following steps: resuscitating cryopreserved Balb / c mouse embryonic stem cells, culturing with a culture solution A containing leukemia inhibition factors, and continuously culturing with a culture solution B containing Activin A after passage; sequentially culturing in an HM culture medium added with specific cell factors, and cooperatively using a cell induction matrix prepared from various components such as hyaluronic acid, 3-aminophenylboronic acid, mono-ammonium glycyrrhizinate and the like. Through the synergistic effect of each culture step and each component in the cell induction matrix, the liver microenvironment is simulated, the cell growth factors are slowly released, and meanwhile, the anti-oxidation characteristic of quercetin is exerted, so that the induced differentiation efficiency of the embryonic stem cells is effectively improved.

The invention provides a non-coding RNA combination for inducing totipotency of mouse embryonic stem cells and application of the non-coding RNA combination, and relates to the technical field of biology. The invention provides a non-coding RNA combination for inducing totipotency of mouse embryonic stem cells. The non-coding RNA combination comprises an enhancer RNA with a sequence as shown in SEQ ID NO: 1, an enhancer RNA with a sequence as shown in SEQ ID NO: 2 and a long-chain non-coding RNA with a sequence as shown in SEQ ID NO: 3. The invention reveals that the CRISPR activation system is utilized to synergistically up-regulate the expression of the specific endogenous non-coding RNA combination for the first time, so that the mouse embryonic stem cells can be efficiently promoted to be converted into bicellular cells. Exogenous transcription factor integration is not needed, so that the safety risk of a genome is avoided; precise targeting is realized through sgRNA, and the defect of non-specific off-target of chemical small molecules is overcome; a remarkable synergistic effect is generated, and the induction efficiency of the two-cell-like cells can be remarkably improved to about 11.6%. The invention provides a safer, more efficient and controllable new strategy for stem cell fate reprogramming and early development research.

The invention relates to the technical field of biological medicines, in particular to a construction method of Gjb2 homozygous mutation deafness mice. Injecting the Gjb2 homozygous mutant mouse embryonic stem cells into a diploid mouse embryo through zona pellucida to prepare a reconstructed embryo; and transferring the reconstructed embryo to a pseudo-pregnant female mouse, so that the postpartum generation mouse is the Gjb2 homozygous mutation deafness mouse. By adopting the construction method disclosed by the invention, the survival ratio of the mouse with homozygous mutation deafness is high, a large number of important mouse models can be provided for follow-up disease treatment exploration, and an important foundation is laid for efficient establishment of mouse models with other different mutations of GJB2.

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.

The application provides an enhancer RNA molecule MZGAe1 and application thereof, and relates to the technical field of biology.The application provides an enhancer RNA molecule MZGAe1, and the nucleotide sequence of the enhancer RNA molecule MZGAe1 is shown in SEQ ID NO:1; meanwhile, the application also provides a specific sgRNA sequence for targeting and activating the molecule and a recombinant carrier thereof; by using the CRISPR activation technology, the expression of the endogenous MZGAe1 in cells can be specifically activated, and the mouse embryonic stem cells can be efficiently promoted to transform into two-cell-like cells. In the application, under the condition of endogenous activation of a single enhancer RNA molecule MZGAe1, the wild-type mouse embryonic stem cells can be efficiently transformed into two-cell-like cells, and the application provides a new efficient and specific research tool and scheme for studying zygote genome activation, early embryonic development events and cell reprogramming.