Assessing miR-451 Activity and Its Role in Erythropoiesis

Methods Mol Biol. 2018:1680:179-190. doi: 10.1007/978-1-4939-7339-2_12.

Abstract

The ability to microinject small RNAs and mRNAs into zebrafish embryos, of different genetic backgrounds, allows for the precise dissection of microRNA processing pathways at the molecular level, while simultaneously provides insight into their physiologic role. Here, we apply such an approach to determine the impact of Argonaute 2 in the processing of miR-451, a vertebrate-specific microRNA required for terminal erythrocyte differentiation. This was achieved using fluorescent microRNA reporter sensor assays and phenotype rescue experiments.

Keywords: Ago2; Erythrocyte; MicroRNA; Microinjection; Zebrafish; miR-451.

MeSH terms

  • Animals
  • Argonaute Proteins / genetics
  • Erythropoiesis / genetics*
  • Gene Expression
  • Gene Expression Regulation*
  • Genes, Reporter
  • MicroRNAs / genetics*
  • Phenotype
  • RNA Interference*
  • RNA, Messenger / genetics
  • Zebrafish
  • Zebrafish Proteins / genetics

Substances

  • Ago2 protein, zebrafish
  • Argonaute Proteins
  • MicroRNAs
  • RNA, Messenger
  • Zebrafish Proteins