A method to delay the aging of mesenchymal stem cells through foxp1 gene editing and mutation
A stem cell and gene editing technology, applied in the field of cell biology, can solve problems such as off-target effects and gene variation, poor miRNA effect, and risks brought by operators, so as to promote proliferation, improve the potential of treating diseases, and improve organ repair Effect
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[0058] This embodiment relates to a method for delaying the aging of human umbilical cord mesenchymal stem cells (MSCs) through FOXP1 gene editing. Using the artificial chromosome episomal-Cas9n vector, two reverse sgRNAs near the FOXP1 gene mutation site were inserted in series; meanwhile, donor DNA containing about 500 bp homology arms at the left and right sides of the mutation site was prepared by PCR. Episomal-Cas9n-FOXP1sgRNA and donor DNA were co-transfected into human umbilical cord mesenchymal stem cells, the cells were diluted and cultured to form single-cell clones, and after antibiotic screening, human-derived FOXP1 (T176G, T277G, K393G) point mutations were identified Umbilical cord mesenchymal stem cells.
[0059] The method comprises the steps of:
[0060] 1) Isolation and culture of human umbilical cord mesenchymal stem cells
[0061] (1) Take out a 15cm dish (petri dish with a diameter of 15cm), cut up the fetal umbilical cord with scissors, suck up and disc...
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