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GONAD: Genome-editing via Oviductal Nucleic Acids Delivery system: a novel microinjection independent genome engineering method in mice.
- Source :
-
Scientific reports [Sci Rep] 2015 Jun 22; Vol. 5, pp. 11406. Date of Electronic Publication: 2015 Jun 22. - Publication Year :
- 2015
-
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 well 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.
- Subjects :
- Animals
CRISPR-Cas Systems genetics
Clustered Regularly Interspaced Short Palindromic Repeats genetics
Elongation Factor 2 Kinase genetics
Embryo, Mammalian cytology
Female
Green Fluorescent Proteins genetics
Hypoxanthine Phosphoribosyltransferase genetics
Mice
Mice, Inbred C57BL
Mice, Knockout
Oviducts cytology
RNA, Messenger genetics
Electroporation methods
Gene Targeting methods
Genetic Engineering methods
Microinjections methods
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 5
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 26096991
- Full Text :
- https://doi.org/10.1038/srep11406