Application of circular RNA to regulation and control of pancreatic islet beta cell proliferation
A technology of beta cells and pancreatic islets, applied in the field of biomedicine, to achieve good application prospects and the effect of promoting proliferation
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Embodiment 1
[0028] In this embodiment, mouse islet β-cell SJb is used as a model for illustration, so a mouse-derived circRNA, ie, circ_mmu_circ_0014773 (SEQ.ID.NO.2), was selected.
[0029] First, it was verified that circ_mmu_circ_0014773 has an endogenously expressed circular RNA sequence in SJb islet β cells. PCR amplification was carried out by designing specific primers for amplifying circular RNA, and the amplified product was sequenced by Sanger to obtain a circular sequence formed by the first link of the linear sequence. figure 1 The middle arrow shows the novel sequence formed by the combination of the last base and the first base of the linear sequence (looping interface sequence).
[0030] Overexpression of circ_mmu_circ_0014773 can effectively promote the proliferation of mouse islet β-cell SJb, and knocking out endogenous circ mmu_circ_0014773 can effectively inhibit the proliferation of mouse islet β-cell SJb. The overexpression and knockout vectors of circ mmu_circ_001477...
Embodiment 2
[0061] Overexpression of circ mmu_circ_0014773 can effectively promote the proliferation of SJb in mouse islet β cells, and knocking out mmu_circ_0014773 can effectively inhibit the proliferation of SJb in mouse islet β cells. The overexpression and knockout of circ_mmu_circ_0014773 were respectively constructed to stably express SJb in mouse islet β cells, and verified The effect of mmu_circ_0014773 on mouse islet β cell SJb proliferation can be further developed as a drug for treating diabetes and has broad application prospects.
[0062] The following experiments were used to verify:
[0063] 1) The effect of overexpression of mmu_circ_0014773 on the survival rate of islet β cells
[0064] Take the overexpressed circular mmu_circ_0014773 (circ_mmu_circ_0014773), overexpressed linear mmu_circ_0014773 (Linear_mmu_circ_0014773), control plasmid (pCDH) cells constructed above, and inoculate them in a 96-well plate, each well containing 100 μl medium, 10 4 The individual cell s...
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