PFD-rAAV-CaMKIIa-DIO-ChrimsonR-tdTomato-WPREs

Promoter Details

CaMKIIa, pyramidal neuron/excitatory neuron promoter, 1293bp.

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.

Virus Type

adeno-associated virus

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.

Regulatory Element Details

DIO: Double-floxed inverse Orientation, is an element that cooperates with Cre recombinase to control the expression of the target gene. If there is no Cre recombinase, the target gene will not be expressed, but in the presence of Cre, the target gene can be expressed normally.

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 freeze and thaw will reduce the titer by 10%. 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.