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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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Cancer Research
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JoVE Journal Cancer Research
A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

DOI:
10.3791/63498-v

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06:48 min

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June 16, 2022

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1Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas,Universidade de São Paulo

Chapters

  • 00:04Introduction
  • 00:52Cell Culture and HuR Overexpression
  • 02:26The Reporter Assay
  • 05:18Results: MicroRNA Biogenesis Reporter Assay Analyzing Maturation in Mammalian Cells
  • 06:17Conclusion

Summary

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We developed an intronic microRNA biogenesis reporter assay to be used in cells in vitro with four plasmids: one with intronic miRNA, one with the target, one to overexpress a regulatory protein, and one for Renilla luciferase. The miRNA was processed and could control luciferase expression by binding to the target sequence.

Tags

Reporter AssayMicroRNA MaturationMammalian CellsRNA-binding ProteinsMicroRNA BiogenesisCell CultureTransfectionLuciferaseHuRMiR-17-92RAB1B3’UTR

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