GONAD: A novel CRISPR/Cas9 genome editing method that does not require ex vivo handling of embryos

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

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Transgenic technologies used for creating a desired genomic change in animals involve three critical steps: isolation of fertilized eggs, microinjection of transgenic DNA into them and their subsequent transfer to recipient females. These ex vivo steps have been widely used for over 3 decades and they were also readily adapted for the latest genome editing technologies such as ZFNs, TALENs, and CRISPR/Cas9 systems. We recently developed a method called GONAD (Genome editing via Oviductal Nucleic Acids Delivery) that does not require all the three critical steps of transgenesis and therefore relieves the bottlenecks of widely used animal transgenic technologies. Here we provide protocols for the GONAD system.

Original languageEnglish (US)
Pages (from-to)15.8.1-15.8.12
JournalCurrent Protocols in Human Genetics
Volume2016
DOIs
StatePublished - Jan 1 2016

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Nucleic Acids
Embryonic Structures
Technology
Gene Transfer Techniques
Genetically Modified Animals
Zygote
Microinjections
DNA
Gene Editing

Keywords

  • CRISPR/Cas9
  • GONAD
  • Gene delivery
  • Genome editing
  • In vivo electroporation
  • Microinjection
  • Transgenic

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

GONAD : A novel CRISPR/Cas9 genome editing method that does not require ex vivo handling of embryos. / Gurumurthy, Channabasavaiah B; Takahashi, Gou; Wada, Kenta; Miura, Hiromi; Sato, Masahiro; Ohtsuka, Masato.

In: Current Protocols in Human Genetics, Vol. 2016, 01.01.2016, p. 15.8.1-15.8.12.

Research output: Contribution to journalArticle

Gurumurthy, Channabasavaiah B ; Takahashi, Gou ; Wada, Kenta ; Miura, Hiromi ; Sato, Masahiro ; Ohtsuka, Masato. / GONAD : A novel CRISPR/Cas9 genome editing method that does not require ex vivo handling of embryos. In: Current Protocols in Human Genetics. 2016 ; Vol. 2016. pp. 15.8.1-15.8.12.
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