Gt(ROSA)26Sor^{em3(CAG-Gf1i,-Atoh1/EGFP,-Pou4f3)Akg}

The targeting vector is designed using a modified Ai3 (Gt(ROSA)26So) vector where the eGFP sequence was replaced with coding regions for growth factor independent 1 transcription repressor (Gfi1), atonal bHLH transcription factor 1 (Atoh1) fused to an enhanced green fluorescent protein (eGFP) and POU domain, class 4, transcription factor 3 (Pou4f3). Specifically, the targeting vector includes (from 5' to 3') a splice acceptor, a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), a loxP-flanked STOP (with stop codons in all 3 reading frames and a triple polyA signal) cassette, Gfi1 sequence, a GSG oligopeptide linker-P2A peptide sequence, Atoh1/eGFP fusion gene, a second GSG oligopeptide linker-P2A peptide sequence, Pou4f3 sequence, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), polyA sequence, followed by an attB/attP-flanked PGK-neo-PGKpA cassette. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. (J:339290)