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

Product Overview

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

Properites

Fluorophore

mCherry

Data

Product Pictures

Publications

Publications (0)

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Customer Reviews and Q&As

Customer Reviews Average Customer Ratings Overall

5.0

Excellent

We used this vector for targeted inhibition in our mouse models

The hEF1a promoter ensured strong and ubiquitous expression across various cell types, and the fDIO system provided precise control over which cells were being manipulated. The fluorescence from the mCherry reporter was bright and easy to detect, which significantly facilitated our analysis.

Excellent

I have utilized the vector in several projects aimed at understanding neural circuitry

The viral vector delivered reliable and high levels of expression in the targeted neurons. The inclusion of the WPRE element seemed to enhance the stability and translation of the transgene, making our results very reproducible. The quality and consistency of this product have been outstanding.

Excellent

This optogenetic tool has greatly facilitated our ability to control and study neuronal activity

The eNpHR 3.0 construct allowed for effective silencing of neural populations, and the mCherry reporter was incredibly helpful for confirming expression patterns. The vector's efficiency in transducing our cells of interest was remarkable, providing us with clear and interpretable data.

Excellent

The overall performance and reliability of this product have been excellent

In our optogenetics studies, the AAV2/9-hEF1a-fDIO-eNpHR 3.0-mCherry-WPRE-pA vector has provided reliable and high-efficiency transduction.

Q&As

How effective is this vector in optogenetic applications?

The AAV2/9-hEF1a-fDIO-eNpHR 3.0-mCherry-WPRE-pA vector is highly effective in optogenetic applications. It allows for precise control over neuronal activity using light. The eNpHR 3.0 component is a third-generation halorhodopsin that efficiently hyperpolarizes neurons in response to yellow light, leading to their inhibition. The mCherry fluorescent marker aids in verifying the expression of the construct in target cells, making it a robust tool for optogenetic studies.

What kind of cells or tissues is this vector compatible with?

The AAV2/9 serotype is well-suited for a wide range of tissues, including CNS neurons and peripheral tissues. It can transduce both dividing and non-dividing cells, making it versatile for various optogenetic research applications. The hEF1a promoter ensures strong and ubiquitous expression across different cell types, enhancing the overall applicability of this vector in neuroscience research.

How do I verify the expression of the eNpHR 3.0-mCherry construct in my cells?

The expression of the eNpHR 3.0-mCherry construct can be verified through several methods. Fluorescence microscopy can be used to detect the mCherry fluorescent marker, confirming the presence of the construct in transduced cells. Additionally, immunohistochemistry or Western blotting using specific antibodies against mCherry or eNpHR can further validate expression. Functional assays involving light stimulation and electrophysiological recording can also confirm the optogenetic functionality of the construct.