Tg(Thy1-COP4*H134R/EYFP)20Gfng

A channelrhodopsin-2 cDNA sequence derived from green alga Chlamydomonas reinhardtii was modified for optimal mammalian expression (beta2 nictinic acytlcholine receptor signal peptide (MAGHSNSMALFSFSLLWLCSGVLGTEF) at the N-terminus, and ER-exporting motif (RSRFVKKDGHCNVQFINVGEKGQRYLA) and Golgi-exporting motif (NSFCYENEVALTSK) (both from potassium channel Kir2.1 gene) at the C-terminus) and modified with a gain-of-function H134R substitution (CAC to CGC) designed to cause larger stationary photocurrents. This B2SP-mhChR2-ER-Golgi sequence (also called B2SP-hChR2-H134R-ER export-Golgi export) was fused in-frame to the amino terminus of an enhanced yellow fluorescent protein sequence (EYFP) via a Not1 site with GCGGCCGCC linker sequence. The resulting mhChR2::YFP fusion protein (also called hChR2-H134R-EYFP) was placed downstream of the modified regulatory region of the "murine thy1.2 gene" (extending from the promoter to the intron following exon 4, excluding exon 3 and its flanking introns), and followed by a bovine growth hormone polyA signal. Mice derived from founder line 20 exhibit high EGFP expression in cortex, CA1, thalamus, superior colliculus, inferior culliculus, brainstem, amygdala and cerebellum. (J:160660)