Article
CRISPR/Cas9-based genome editing in mice by single plasmid injection.
Methods in enzymology - 1 Jan 2014
Fujihara Yoshitaka, Ikawa Masahito
Abstract excerpt
CRISPR/Cas-mediated genome modification has opened a new era for elucidating gene function. Gene knockout mice can be generated by injecting humanized Cas9 (hCas9) mRNA and guide RNA (sgRNA) into fertilized eggs. However, delivery of RNA instead of DNA to the fertilized oocyte requires extra preparation and extra care with storage. To simplify the method of delivery, we injected the circular pX330 plasmids...
Topics
- Animals
- CRISPR-Associated Proteins
- Clustered Regularly Interspaced Short Palindromic Repeats
- Endonucleases
- Gene Targeting
- Green Fluorescent Proteins
- HEK293 Cells
- Humans
- Mice
- Microinjections
- Mutagenesis