Tissue-specific RNA interference in post-implantation mouse embryos using directional electroporation and whole embryo culture

Federico Calegari, Anne Marie Marzesco, Ralf Kittler, Frank Buchholz, Wieland B. Huttner

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

In mammals, embryonic development is more difficult to analyze than in non-mammalian species because this development occurs in utero. Interestingly, whole embryo culture allows the normal development of mouse post-implantation embryos for up to 2 days in vitro. One limitation of this technology has been the difficulty of performing loss-of-gene function studies in this system. RNA interference (RNAi), whereby double-stranded RNA molecules suppress the expression of complementary genes, has rapidly become a widely used tool for gene function analyses. We have combined the technologies of mouse whole embryo culture and RNAi to allow the molecular dissection of developmental processes. Here, we review the manipulation by topical injection followed by directional electroporation of endoribonuclease-prepared siRNA to demonstrate that this technology may be useful to knock down genes in a tissue- and region-specific manner in several organs of the developing mouse embryo.

Original languageEnglish (US)
Pages (from-to)92-102
Number of pages11
JournalDifferentiation
Volume72
Issue number2-3
DOIs
StatePublished - Mar 2004

Fingerprint

Electroporation
RNA Interference
Embryonic Structures
Technology
Embryonic Development
Gene Knock-In Techniques
Endoribonucleases
Double-Stranded RNA
Small Interfering RNA
Genes
Dissection
Mammals
Gene Expression
Injections

Keywords

  • Endoribonuclease-prepared short interfering RNAs
  • In vivo electroporation
  • RNA interference
  • Whole embryo culture

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Developmental Biology
  • Cell Biology

Cite this

Tissue-specific RNA interference in post-implantation mouse embryos using directional electroporation and whole embryo culture. / Calegari, Federico; Marzesco, Anne Marie; Kittler, Ralf; Buchholz, Frank; Huttner, Wieland B.

In: Differentiation, Vol. 72, No. 2-3, 03.2004, p. 92-102.

Research output: Contribution to journalArticle

Calegari, Federico ; Marzesco, Anne Marie ; Kittler, Ralf ; Buchholz, Frank ; Huttner, Wieland B. / Tissue-specific RNA interference in post-implantation mouse embryos using directional electroporation and whole embryo culture. In: Differentiation. 2004 ; Vol. 72, No. 2-3. pp. 92-102.
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