Not applicable

The region deleted via CRISPR-Cas9 corresponds to a transgenetically verified enhancer. Deletion of the enhancer results in homozygote mutants losing Nr2f1 expression in the ventricle zone in rostral regions of the dorsomedial pallium (cortex) at embryonic day 12.5, and there was a mild rostral to caudal expansion of somatosensory neocortex at postnatal day 5.

Four hs1172 sgRNAs were used to delete hs1172 in C57BL/6 mice. The Cas9 gene was PCR amplified and the resulting T7-promoter-Cas9 DNA fragments were gel excised, purified, and in vitro transcribed. sgRNAs were assembled by oligo annealing/extension and then in vitro transcribed. RNA was purified and prepared for injection by mixing Cas9 mRNA, and the three sgRNAs in microinjection buffer. E0.5 embryos were injected. Enhancer deletion mice were genotyped by PCR. The mice were outbred with wild type C57BL/6 for three generations to reduce the number of potential off-target mutations.

wild-type

The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact mmrrc@ucdavis.edu. Older strains may not have this information.

John Rubenstein, M.D., University of California, San Francisco.

In preparation, submitted, or in press