AAV2/9-hEF1a-fDIO-eNpHR 3.0-mCherry-WPRE-pA

Description

fDIO, or fDiO (Flp controlled Double-floxed inverse Orientation), is an element that cooperates with Flp to control the expression of the target gene. It is generally composed of two pairs of mutually incompatible Frt sequences. The characteristic sequence is Frt-interval 1-Frt5-GOIr -Ftrr-Interval 2-Frt5r. When Flp is present, any pair of Frt sequences may recombine. Because each pair of Frt is in the opposite direction, recombination will cause the sequence between them to be reversed. For example, Frt recombines first to form Frt-GOI- Frt5r-interval 1r-Frtr-interval 2-Frt5r, then the direction of the two Frt5 is changed from the previous opposite to the same, the second recombination between them will make Ftrr missing, forming Ftr-GOI-Frt5r, due to Frt No recombination can occur between Frt5 and Frt5, and the position of the target gene is fixed. Frt5 reorganizes first, and the result is the same. Usually we insert the target gene so that its sequence is opposite to the promoter, that is, it is not expressed at the beginning, and the target gene begins to be expressed after Cre acts.

The hEF1a promoter, cloned from the promoter region of human translation elongation factor 1A1 (eukaryotic translation elongation factor 1 alpha 1 (EEF1A1)), is 1.3 kb in length, expresses constitutively, and has no cell tissue specificity.

NpHR, a light-driven chloride ion pump, belongs to Halorhodopsin. The common version is NpHR3.0, which adds the endoplasmic reticulum forward transport signal and the Golgi transport signal to the cell membrane on the basis of NpHR, which improves its expression on the cell membrane and is the most commonly used photoinhibition tool. NpHR is usually driven by yellow light around 570nm and can be used simultaneously with hChR2 (H134R).

Tracer Type

Optogenetics

Functional Gene

NpHR

Activity

Inhibition

Serotype

AAV9

Virus Type

Adeno-Associated Virus (AAV)

Applications

Optogenetics

Research Areas

Neural Circuit Mapping

Promoter

EF1a

Expression

Flp-dependent

Fluorophore

mCherry