GONAD

Genome-editing via Oviductal Nucleic Acids Delivery system: A novel microinjection independent genome engineering method in mice

Gou Takahashi, Channabasavaiah B Gurumurthy, Kenta Wada, Hiromi Miura, Masahiro Sato, Masato Ohtsuka

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Microinjection is considered the gold standard technique for delivery of nucleic acids (NAs; transgenes or genome editing tools such as CRISPR/Cas9 systems) into embryos, for creating genetically modified organisms. It requires sophisticated equipment as wel as well-trained and highly skilled personnel to perform the micro-injection technique. Here, we describe a novel and simple microinjection-independent technique, called Genome-editing via Oviductal Nucleic Acids Delivery (GONAD). Using GONAD, we show that NAs (e.g., eGFP mRNA or Cas9 mRNA/sgRNAs) can be effectively delivered to pre-implantation embryos within the intact mouse oviduct by a simple electroporation method, and result in the desired genetic modification in the embryos. Thus GONAD can bypass many complex steps in transgenic technology such as isolation of zygotes, microinjection of NAs into them, and their subsequent transfer to pseudo-pregnant animals. Furthermore, this method can potentially be used for genome editing in species other than mice.

Original languageEnglish (US)
Article number11406
JournalScientific Reports
Volume5
DOIs
StatePublished - Jun 22 2015

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Microinjections
Nucleic Acids
Genome
Embryonic Structures
Clustered Regularly Interspaced Short Palindromic Repeats
Genetically Modified Organisms
Messenger RNA
Electroporation
Oviducts
Zygote
Transgenes
Gene Editing
Technology
Equipment and Supplies
Injections

ASJC Scopus subject areas

  • General

Cite this

GONAD : Genome-editing via Oviductal Nucleic Acids Delivery system: A novel microinjection independent genome engineering method in mice. / Takahashi, Gou; Gurumurthy, Channabasavaiah B; Wada, Kenta; Miura, Hiromi; Sato, Masahiro; Ohtsuka, Masato.

In: Scientific Reports, Vol. 5, 11406, 22.06.2015.

Research output: Contribution to journalArticle

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