Agtr1a^em1(cre)Zak

The bicistronic Agtr1a-2A-Cre knock-in allele was created using homologous recombination (aided by targeted CRISPR/cas9 endonuclease activity) to insert a viral 2A oligopeptide sequence (T2A; mediates ribosomal skipping) and a Cre recombinase gene (cre) immediately upstream of the endogenous STOP codon of Agtr1a gene. The targeting vector, Cas9 endonuclease and single guide RNA were mixed and injected into the pronuclei of FVB/N fertilized zygotes. Embryos were transferred to pseudopregnant females, and correctly targeted pups were maintained on mixed FVB/C57BL/6J background. Two independent knock-in lines were crossed to reporter mice, and reporter expression patterns from these lines were identical. (J:256118)