rAAV-CAG-ChrimsonR-P2A-tdtomato-WPRES

Promoter Details

CAG, strong promoter, is an artificially constructed combined promoter, consisting of cytomegalovirus early enhancer and chicken β-actin promoter, 1731bp.

Functional Gene Details

ChrimsonR: A new type of light-controlled ion channel that can respond to red light around 600nm was discovered through high-throughput sequencing in the Boyden laboratory. A point mutation of K176R was made on the basis of Chrimson (CnChR1), which speeds up the channel closing speed, making it possible to effectively respond to light signals at least 20Hz and below. When used with Chronos, it is possible to control two different neurons with two fluorophores. The excitation light wavelength is 590nm.

Fluorescent Tag

tdtomato

Fluorescent Protein Details

tdTomato: Red fluorescent protein, excitation wavelength 554nm, emission wavelength 581nm. 2A: The 2A sequence is a short peptide sequence. When the ribosome translates it, it will automatically jump away and continue to translate the downstream reading frame, thereby translating the same reading frame into two independent proteins. Used in non-fusion expressions. Commonly used 2A sequences include T2A and P2A.

tdTomato: Red fluorescent protein, excitation wavelength 554nm, emission wavelength 581nm.

Application

Expressing light-sensitive ion channel proteins, such as ChR2, ChrimsonR, etc., in neurons through rAAV viral vectors can specifically excite neurons and explore the effects of light stimulation on neurons, neural circuits and animal behavior.

Notes

1. Short-term storage at 4°C; long-term storage at -80°C

2. It can be stored at -80°C for more than 6 months. Each time it is frozen and thawed, the titer will be reduced by 10%, and repeated freezing and thawing should be avoided; if the virus is stored for more than 6 months, it is recommended to re-measure the virus titer before use

3. According to the experimental needs, the virus can be appropriately diluted and divided into the volume required for each experiment.