Assaying NanoLuc Luciferase Activity from mRNA-Injected Xenopus Embryos

Methods Mol Biol. 2019:1920:33-39. doi: 10.1007/978-1-4939-9009-2_3.

Abstract

The earliest steps of animal development depend upon posttranscriptional events that drive the embryonic cell cycle and guide cell fate decisions. The analysis of post-transcriptional regulatory events has relied upon the use of chimeric reporter mRNAs that encode firefly luciferase fused to potential regulatory sequences. A new and more sensitive luciferase developed recently called NanoLuc has the potential to improve reporter studies and provide new insights into the regulation of embryonic processes. Here I describe how to create and analyze reporter mRNAs encoding NanoLuc luciferase using extracts from microinjected Xenopus embryos.

Keywords: Luciferase; NanoLuc; Xenopus embryos; mRNA microinjection.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Embryo, Nonmammalian
  • Gene Expression*
  • Genes, Reporter*
  • In Vitro Techniques
  • Luciferases / genetics*
  • Luminescent Measurements* / methods
  • Microinjections*
  • Plasmids / genetics
  • RNA, Messenger / administration & dosage
  • RNA, Messenger / genetics*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transcription, Genetic
  • Xenopus laevis / genetics*
  • Xenopus laevis / metabolism

Substances

  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Luciferases