CRISPR/Cas9-based genome editing in mice by single plasmid injection
Y Fujihara, M Ikawa - Methods in enzymology, 2014 - Elsevier
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 expressing both
hCas9 and sgRNA and found that mutant mice were generated as efficiently as with RNA …
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 expressing both
hCas9 and sgRNA and found that mutant mice were generated as efficiently as with RNA …