Tg(Pvalb-COP4*H134R/EYFP)15Gfng

First, a channelrhodopsin-2 cDNA sequence derived from green alga Chlamydomonas reinhardtii was modified to harbor condons optimized for mammalian expression and a gain-of-function H134R substitution (CAC to CGC) designed to cause larger stationary photocurrents. This mhChR2 sequence (also called hChR2-H134R) was fused in-frame to the amino terminus of an enhanced yellow fluorescent protein sequence (EYFP) via a Not1 site with GCGGCCGCC linker sequence. This mhChR2::YFP fusion protein (also called hChR2-H134R-EYFP) sequence was inserted into the coding region of the parvalbumin (Pvalb or Prv) locus on the RP23-305H12 bacterial artificial chromosome (BAC) via homologous recombination. Mice from line 15 exhibit high EGFP expression in cerebellum, medium EGFP expression in reticular thalamic nuclei, and low EGFP expression in brainstem. (J:160657)