Gt(ROSA)26Sor^{em3(CAG-Brainbow2.1,-Pik3ca*)Koo}

CRISPR-targeting inserted the Red2Onco construct, containing PIK3CAH1047R (addgene 12524), into the tdimer2 reporter within the Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle. The Brainbow2.1 (also known as Confetti) reporter construct is a modification of the Brainbow reporter with a strong CAGG promoter, a loxP site, a PGK-Neo-pA cassette (serving as a transcriptional roadblock), and coding region composed of two adjacent floxed head-to-tail tandem dimers. The first head-to-tail dimer contains a loxP site and humanized Renilla GFP (hrGFPII; with nuclear localization signal plus polyA sequence) in forward orientation and a loxP site and monomeric EYFP (mYFPA206K plus polyA sequence) in reverse orientation. The second head-to-tail dimer contains a loxP site and tdimer2 RFP plus polyA sequence in forward orientation, and a loxP site and mCerulean CFP (with membrane tethering palmitoylation sequence plus polyA sequence) in reverse orientation. A single frt site is located at the 3' end of the reporter construct. (J:322623)