Gt(ROSA)26Sor^{em1(CAG-Mapk1*/mTagBFP2,-H2B/iRFP713,-NFATC2*/mRuby3)Hyku}

CRISPR/cas9 genome editing technology is used to insert a construct containing from 5’ to 3’: a CAG promoter, a loxP-neomycin cassette-loxP (LSL) cassette followed by an Erk (formally, Mapk1) kinase translocation reporter (KTR) fused to the mTagBFP2 reporter, a tP2AT2A sequence, a human histone 2B chromatin/nucleus tag fused to the iRFP713 reporter, an IRES, and the N terminal regulatory domain (residues 1-399) of mouse Nfatc2 fused to the mRuby3 reporter inserted after exon 1 of the Gt(ROSA)26Sor locus. The ERK-KTR sensor consists of an ERK1/2 binding site and phosphorylation motif adjacent to phosphorylation activated nuclear export signal (NES) and nuclear localization sequence (NLS) fused to a fluorescent protein (mTagBFP2). (J:342890)