5 results about "PDX1" patented technology

Methods are provided for generating pancreatic endocrine cells from pluripotent stem cells (PSCs) using epigenetic modifiers. A fusion protein comprising catalytically inactivated Cas9 (dCas9) and an embryonic ectoderm development (EED)-binding domain (EBdCas9) is used with guide RNAs (gRNAs) to target regulatory regions of transcriptional regulators including EOMES, PDX1, and NGN3. Sequential targeting of PDX1 and NGN3 promoters with EBdCas9 removes repressive H3K27me3 marks, increases activating H3K27ac modifications, and accelerates differentiation into insulin-producing β-cells. The methods significantly increase β-cell yields (~4-6 fold), reduce non-pancreatic lineage contaminants (CDX2+, SOX2+, enteroendocrine cells), enhance mitochondrial function, and produce glucose-responsive cells with superior functional maturation in vivo. The epigenetic interventions enforce pancreatic developmental fates while activating NGN3-dependent transcriptional programs necessary for endocrine cell differentiation, insulin secretion, and metabolic maturation. Cells and compositions produced by these methods are also provided.

The invention discloses a pancreatic tissue specific expression recombinant adeno-associated virus vector and application thereof, and belongs to the technical field of biological medicine. Comprising the following steps: S1, cloning a pancreatic specific promoter: jointly using Ins2 and Pdx1 promoters to prepare a Pdx-1 / Ins2 promoter; performing serotype and capsid mutation optimization; s2, insertion of an exogenous gene expression cassette: cloning a target gene to the downstream of a Pdx-1 / Ins2 promoter to construct a'promoter-exogenous gene-polyadenylic acid signal 'expression unit; s3, regulation and control element integration: inserting an HS4 insulator sequence into two sides of an expression box, and adopting Gibson assembly to realize seamless connection; constructing an SIN structure: deleting a U5 region in ITR through site-specific mutagenesis or enzyme digestion, and verifying that no residual promoter activity exists; s4, virus packaging and purification: packaging rAAV particles by adopting an HEK293 cell three-plasmid co-transfection system; the three plasmids comprise a carrier plasmid, an auxiliary plasmid and a packaging plasmid; and determining the purity after purification.

The invention discloses a pancreatic beta cell precursor cell transplantation chip based on human-derived iPSC (induced pluripotent stem cell) as well as a preparation method and application of the pancreatic beta cell precursor cell transplantation chip. The chip is composed of beta cell precursor cells (expressing Pdx1, Nkx6.1 and NeuroD1) and a porous degradable bracket (PCL / hyaluronic acid composite material), wherein the beta cell precursor cells (expressing Pdx1, Nkx6.1 and NeuroD1) are obtained by performing four-stage induction on autologous iPSC. The preparation method comprises the steps of autologous iPSC acquisition, beta cell precursor cell induction, three-dimensional bracket preparation and chip assembly. After the chip is implanted into the pancreas of a patient through minimally invasive surgery, precursor cells are differentiated into mature beta cells in vivo, insulin is secreted, the stent is gradually degraded, the pancreas islet function can be fundamentally improved, and the chip is used for treating type I and type II diabetes mellitus. The invention avoids immunological rejection, improves cell survival and functional stability, and has significant clinical transformation value.

The application discloses application of MST1 interfering RNA in promoting in-vitro differentiation of ESCs into insulin-secreting cells, and belongs to the technical field of biotechnology. In the process of in-vitro directional differentiation of rat ESCs into beta cells, the application infects by using an MST1 shRNA lentivirus vector, and indexes are detected and functions are determined by using cell biology and molecular biology methods, and it is found for the first time that the efficiency of inducing ESCs to differentiate into beta cell-like cells can be improved by reducing the expression of MST1. Meanwhile, how MST1 realizes the regulation of beta cell maturation by means of PDX1 in the process of directional differentiation of rat ESCs into beta cells is described, the reason affecting in-vitro maturation of beta cells is discussed, and thus the system of directional differentiation of ESCs into mature beta cells is optimized, which has important significance for obtaining beta cells with normal insulin secretion function in-vitro.

The present invention relates to a peptide having anti-obesity and anti-diabetes activities and uses thereof. The peptide of the present invention increases the expression of insulin genes and genes of PDX1 and GLP1R involved in insulin secretion, increases the secretion of insulin itself, and increases the expression levels of phospho-CREB and POMC factors associated with satiety signaling. In addition, the peptide of the present invention exhibits anti-obesity activity. Therefore, the peptide of the present invention can be used for treating, preventing, and alleviating obesity or diabetes, and can be advantageously used for lowering blood glucose levels of a person in need of blood glucose control.