Wednesday, November 27, 2013

Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA - Ikawa lab papers, November 2013.

The first of these 2 papers is the first report of mouse mutagenesis by pronuclear injection of circular plasmids expressing CRISPR gRNA and Cas9 mRNA.    In a follow-up paper, the authors demonstrate this approach on many more genes; their approach is to validate the CRISPR plasmids in cell transfections first, then use the effective ones for zygote injections.

The plasmid used in both papers was PX330, which was made by the Zhang lab - links for this are at the bottom.   

1.  Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA.

Daisuke Mashiko, Yoshitaka Fujihara, Yuhkoh Satouh, Haruhiko Miyata, Ayako Isotani & Masahito Ikawa

Scientific Reports
 
3,
 
Article number:
 
3355
 
doi:10.1038/srep03355
Received
 
3 Accepted 12 November 2013 Published
Feasibility for a large scale mouse mutagenesis by injecting CRISPR/Cas plasmid into zygotes.
Mashiko D, Young SA, Muto M, Kato H, Nozawa K, Ogawa M, Noda T, Kim YJ, Satouh Y, Fujihara Y, Ikawa M.
 2014 Jan;56(1):122-9. doi: 10.1111/dgd.12113. Epub 2013 Dec 26.

Links for PX330 and PX335:  



PX335 is similar to PX330 except PX335 has a nickase form of Cas9.

No comments:

Post a Comment