Microinjection of oocytes and embryos with synthetic mRNA encoding molecular probes

Methods Cell Biol. 2019:150:189-222. doi: 10.1016/bs.mcb.2018.10.012. Epub 2018 Dec 21.

Abstract

We describe methods and techniques for introduction of molecular probes in the form of synthetic mRNA by rapid repetitive microinjection into oocytes or early embryos of echinoderms and various invertebrates. Construct assembly is followed by standard kit-based in vitro mRNA synthesis, with slight modifications to optimize expression and clean-up. Variations of a basic microinjection procedures are detailed for echinoderms: starfish oocytes (Patiria miniata or other species), purple urchin (Strongylocentrotus purpuratus) and sand dollar (Dendraster excentricus) zygotes, with notes included for other invertebrate eggs and embryos as well.

Keywords: Echinoderm; Embryo; Fluorescent protein; Microinjection; Oocyte; Synthetic mRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Review

MeSH terms

  • Animals
  • Embryo, Nonmammalian / cytology*
  • Embryo, Nonmammalian / physiology*
  • Microinjections / methods*
  • Molecular Probes / administration & dosage*
  • Oocytes / cytology*
  • RNA, Messenger / administration & dosage*
  • Sea Urchins / genetics
  • Starfish / genetics

Substances

  • Molecular Probes
  • RNA, Messenger