Anion-conducting channelrhodopsins with tuned spectra and modified kinetics engineered for optogenetic manipulation of behavior
| Products/Services Used |
Details |
Operation |
| PCR Cloning and Subcloning |
Briefly, ChR cDNAs were cloned into p-EGFP-C1 vectors using NheI and AgeI restriction sites. EGFP was replaced by mCherry using AgeI and XhoI restriction sites except for iChloC variants where a p-EGFP-N1 vector was used. The QuikChange II kit (Agilent Technologies, Santa Clara, CA) was used to exchange single or multiple amino acids (C128A, T159G and G163A) in ChloC/iChloC based variants. For the iC++ based approach, ChR variants with the replaced N-terminus of iC++ and all iC++ homologous mutations were constructed in-silico, synthesized (GenScript, NJ) and cloned into p-EGFP-C1 vectors as described above. |
Get A Quote |
