Аннотация:It has been successfully demonstrated to edit target gene (knock-in of reporters or disruption of genes) by recently developed technologies, such as Zinc Finger Nucleases of artificial nuclease (ZFNs) or Transcription Activator-Like Effector Nucleases (TALENs).More recently, efficient genome editing has been reported by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system in variety of vertebrates as well as in mice, which would be more broadly used in the future.In the animal facility of the Institute for Virus Research, Kyoto University, we are generating CRISPR/Cas9 mediated mutant mice by two different methods.First one is by Wang et al. reported in 2013, RNA is injected into the cytoplasm of mouse pronuclear stage fertilized eggs.Another one is by Ikawa et al. of Research Institute for Microbial Diseases, Osaka University, a plasmid is injected into the pronucleus.Although we observed no obvious difference in efficiency or reliability between these two methods at the moment, development of CRISPR/Cas9 system would be more important for genome editing such as generation of knock-in or knock-out mice.Here we would report a short summary of our experiments together with some examples of genome-edited mutant mice.
Год издания:2014
Источник:EXPERIMENTAL ANIMALS
Ключевые слова:Science, Research, and Medicine, Pluripotent Stem Cells Research
