CRISPR/Cas9 allows scientists to develop gene-targeted mice more quickly and for less cost than traditional methods. CRISPR/Cas can gather mutations from the injection of fertilized eggs, which eliminates the need for ES cells and targeting vectors, among other protocols. (Sander JD, Joung JK. 2014. CRISPR-Cas systems for editing, regulation and targeting genomes. Nature Biotechnology 32(4):347-355)
Fertilized eggs will be obtained by mating (C57BL/6 X C3H)F1 or (C57BL/6 X SJL)F1 females to (C57BL/6 X C3H) F1 or C57BL/6 X SJL)F1 males, respectively. Eggs obtained from other strains of mice are available upon consultation with the Vivarium and Transgenics Core. The fertilized eggs will be injected and then incubated overnight to assess the construct’s toxicity. Eggs dividing to the two-cell stage will be implanted in pseudopregnant foster mothers.
RNA injection is prioritized by the RNA sample’s receipt date; however, clients submitting multiple samples will have one construct injected per week to prevent a backup of samples from other clients.